JP2001500964A - Treatment of the eye using a synthetic thyroid hormone composition - Google Patents

Abstract

  (57) [Summary] The present invention includes the discovery that synthetic thyroid hormone can be used as a composition for reducing intraocular pressure in vivo. Methods for screening synthetic thyroid hormones for effects on intraocular pressure, aqueous pressure, hydraulic conductivity, hyaluronic acid secretion, and extracellular matrix assembly are provided. Also provided are methods of treating excessive intraocular pressure with synthetic thyroid hormone and compositions and methods of treating glaucoma.

Description

DETAILED DESCRIPTION OF THE INVENTION Treatment of the eye using a synthetic thyroid hormone composition                                 Introduction Technical field   The present invention relates to a composition for the treatment of synthetic thyroid hormone, elevated intraocular pressure and glaucoma. Articles and methods. Background   Glaucoma afflicts about 2 million people in the United States and about 145 million worldwide . About 2% of the population over the age of 35 suffer from some form of glaucoma, and It is responsible for about 12% of all cases of Ming. Suffers from glaucoma despite its easy diagnosis Therapy to reduce intraocular pressure (IOP) in patients is often Often not inappropriate. Topical and oral medications for reducing intraocular pressure The use of is often limited by the side effects of those medicaments. In other cases In some cases, the use of medical procedures to treat glaucoma may result in progressive damage to the optic nerve. It has not been successful in lowering intraocular pressure sufficiently to prevent it.   Accordingly, a method for the treatment of glaucoma and other medical conditions of the eye or epithelium, There is a need for compounds and compositions. SUMMARY OF THE INVENTION   Prior to the invention described herein, systemic levels of thyroid hormone The change in the IOP due to the change has been presumed to be indirect or Thyroid hormone-induced alterations in IOP Is mediated through adrenergic α or β receptors located in the eye. Was thought to be. Aspects of the present invention now relate to thyroid formol in ocular physiology. He first recognized the important role of the receptor. Thyroid receptors in the eye Provides the opportunity to directly mediate thyroid hormone effects in cells. In particular, The thyroid receptor of trabecular meshwork (“HTM”) cells is Potentially mediates the amount of extracellular hyaluronic acid ("HA") maintained by vesicles Which, in turn, can affect the amount of aqueous humor outflow.   The present invention relates to cells containing thyroid hormone receptors and cells related to GAG production. Vesicle process mediates glycosaminoglycan ("GAG") production In vitro or to identify compounds that are useful as therapeutics, such as compounds I understand clearly that it will be a useful component in vivo. Therapeutic (th Methods to identify compounds useful as erapeutics are: 1) cells that secrete GAGs, For example, a certain compound is brought into contact with trabecular meshwork cells, and Detecting the binding of the compound to the cells when the concentration is less than or equal to the molar concentration. including.   Furthermore, the present invention relates to thyroid hormones and synthetic thyroid hormones (synthetic thyr hormones). oid hormones (SHTs)) regulate GAG production or HA secretion from cells such as trabecular meshwork cells. Understand clearly that it can be mediated directly. Therefore, the present invention The method comprises reducing the amount or activity of a substance or cellular process in a desired cell or tissue. The compounds, particularly STHs, are used to modulate GAG production or HA secretion. Preferably, such cells are trabecular meshwork cells, ciliary cells of the eye, Endothelial cells of the eye, endothelial cells other than the eye, and skin and internal organs Organ fibroblasts. Most In such cases, non-systemic application of STH is preferred. However, STHs may be administered systemically it can.   The present invention provides an ophthalmologically effective amount of synthetic thyroid hormone for the eye in need of treatment. Also provided is a method of treating glaucoma comprising administering to the eye. Dosing mode is local , Intraocular implantation or injection, and systemic. Preferably, the use of STH for the eye Administration can be topical using an ophthalmically acceptable pharmaceutical carrier or implantable or surgically implantable. Is a non-systemic administration via an injectable device, which is responsible for maintaining STH in the eye. Biodegradable or non-biodegradable polymers that can be released continuously Can be included. Eye inserts containing STH in a sustained release polymer have also been used Can be BRIEF DESCRIPTION OF THE FIGURES   FIG._Three-Supplement or T_Three-Removal (T_Three-Stripped) medium (N_oT_x) In culture Figure 4 shows a graph comparing HA production by isolated HTM cells. By the second day, T_Three−Supplement There was a 57% relative decrease in HA secretion from the cells. By the fourth day, this difference It increased to 66%.   FIG. 2 shows T cells for HA production by cultured HTM cells._ThreeShows the effect of concentration. HA student Maximum inhibition of fertility is 10^-7~Ten^-6T between M_ThreeIt occurred in.   FIG.^-7Incubated for 4 days with or without M T3 4 is a chart showing the hydraulic conductivity of exposed HTM cells. . Control cells not treated with T3 were treated with an appropriate amount of ethanol medium.   FIG.^-7Incubated for 8 days with or without M T3 4 is a chart showing hydraulic conductivity of HTM cells obtained.   5 to 9 show the structures of various STHs. Description of specific aspects Abbreviation   HA represents hyaluronic acid.   GAG represents glucosaminoglycan.   HTM stands for human trabecular meshwork.   IOP represents intraocular pressure.   STH stands for synthetic thyroid hormone. Definition   Intraocular pressure ("IOP") is typically measured in the eye, especially in the anterior chamber. Refers to the pressure associated with the aqueous components within. Intraocular pressure is onometry). In the human non-glaucoma population, IOP Seems to be normally distributed. IOP average as measured by applanation intraocular Is 15.4 (σ ± 2.5^*) MmHg (sitting down The IOP average value is 16.1 (σ ± 2.8) mmHg as measured by Non-glaucoma patients The person's IOP value is only an approximation. The actual frequency distribution of intraocular pressure in this population is Higher levels due to different subpopulations within the general population (eg, glaucoma and age) Can be distorted.   Glaucoma typically causes impaired vision or damage to the optic nerve. Refers to the IOP of the eye of the individual producing the wound. As a general guide, 21mmHg (average +2 IOP above σ) should occur in less than 2.5% of the general population, and Pressures of more than 24 mmHg (mean + 3σ) should occur within less than 0.15% of the 21mm IOPs above Hg indicate glaucoma development, and IOPs above 24 mmHg indicate glaucoma It is a strong indicator. Patient's eyes have visual impairment or elevation over time Increased IOP as it changes in their susceptibility to optic nerve damage from IOP The value must be considered as an indicator rather than a diagnosis of glaucoma. Field of view Evaluation of the optic nerve also ensures that patients do not have abnormally high IOP. Is preferably evaluated. Glaucoma is a closed angle glaucoma ) And open angle glaucoma. Angle-closure glaucoma with rainbow The apposition of the trabecular meshwork to the color. Open-angle glaucoma can occur in the trabecular meshwork or Resistance to aqueous humor outflow that exceeds this. Glaucoma is the optic nerve in normal IOP It includes references to low tension glaucoma, which refers to damage.   Synthetic thyroid hormone (“STH”) binds to thyroid receptors and is normal With partial thyroid receptor hormone function, partial agonist or A molecule that acts as an antagonist. An agonist is a compound whose receptor A molecule that mimics the action of a natural molecule when bound to a molecule. Usually, STHs are T_ThreeMachine Agonists or partial agonists, and using chemical synthetic pathways Will be made. STHs are usually found in T cells isolated from natural tissues._ThreeAnd T_FourDoes not contain . Because the chemically synthesized T_ThreeAnd T_FourPureer preparations are readily available Because it can be Reduces secretion of hyaluronic acid ("HA") from cells STHs or new STHs known in the art that are particularly useful in HA secretion in the eye Regular STHs are further described herein. Preferably, STH is a thyroid hormone 10 μM or less compared to HA secretion in the absence or thyroid hormone-free medium Will decrease HA secretion from human trabecular meshwork cells cultured at a concentration of. STH is S Including ester derivatives of THs.   For purposes of the present invention, the term “thyroid gland”, a term used interchangeably herein, is used. Remon ”or“ thyroid hormone-like compound ” Pure or substantially pure natural or recombinant thyroid hormone containing a gand binding domain Including thyroid hormone receptors such as mon alpha or beta receptors or rat nuclei Using the preparations, (Lavin, T.N. Mechanisms of Thyroid Hormone action . In the textbook of Endocrinology (De Groot, Ed.), 2nd Editiom, W. B. Sa unders, pub. (1989) and Aprileti J. et al. et al J .; Biol. Chem. 263 p. 9409-9 417, 1988) (when tested in a receptor binding assay) Thyroid with chemical affinity constant, KD, less than 1 μN A peptide that binds at least partially to the hormone receptor TR-α or β. Any chemical entity. Such ligands are those whose natural Can be considered a hormone if they have the same agonistic effect as a hormone Or when those compounds antagonize the effects of their natural hormones Can be considered. Partial agonists / antagonists also exist Can be. (Suitable ligands can be agonists or antagonists. Wear. )   Secretion, when used in the context of describing cellular processes, typically refers to A cellular process that transports molecules from the inside of a vesicle to an extracellular location. Glycosaminoguri In the case of cans ("GAG"), secretion is achieved through a number of cellular processes, including post-translational processing. And delivery to appropriate cell sites for transport from cells and cells. Will be. However, secretion refers to the net transport of molecules, and not necessarily It does not refer to a particular step in the secretory process of the molecule.   Synthesis, when used in the context of describing cellular processes, typically involves synthesis. Refers to cellular processes involved in the production of cells, such as GAGs, such as HA. Synthesis made molecule The term synthesis is used herein as it can include the transport of that molecule from the producing cell. The term secretion described in the book Including references.   Manufacturing, when used in the context of describing cellular processes, typically involves Cellular processes involved in maintaining a steady state level of a child, eg, GAG, eg, HA Say. Thus, manufacturing refers to the cellular processes involved in synthesis and secretion. Manufacture the molecule Maintenance of steady state levels, degradation pathways, and separation into cells or in the extracellular matrix. It also refers to cells and extracellular processes that describe extracellular structural elements that anchor the offspring.   An ophthalmologically effective amount is typically the amount of such therapeutic agent required for such treatment. When delivered to a complex eye tissue in a combination and in a corresponding dosing technique A therapeutic agent sufficient to reduce intraocular IOP or prevent elevation of IOP, e.g., STH Means the amount of Typically, elevated IOP is associated with ocular hypertension (without optic nerve damage, Elevated IOP), primary glaucoma, secondary glaucoma (eg, systemic steroid treatment) or It may be associated with hypothyroidism. Described in this specification STHs may be used to treat elevated IOP associated with the above medical conditions Can be. Preferably, an ophthalmologically effective amount prevents optic nerve damage or vision impairment To reduce the IOP sufficiently to make At least 10%, preferably at least 30%, of elevated IOP levels And most preferably at least 50%. The expected normal IOP is typically , (When measured by flat pressure tonometer, sitting position), less than 22 mmHg, for example, less than 15 mmHg Will. Expected normal IOP is low enough to prevent optic nerve damage Should. The success of an STH procedure for reducing IOP has been measured using other formats. You can also. For example, a patient may have an elevated IOP of 34 mmHg and a target IOP of 17 mmHg. Can be included. Preventing optic nerve damage for the patient is 50% in IOP to achieve target IOP Would need to be reduced. This percentage drop in the IOP And the following formula: Can be described more simply. The IOP measurement is based on the ophthalmologically effective Monitor ophthalmic STH procedures to tailor different patient responsiveness to procedures And methods for varying dosages are described more fully herein. Will be revealed. Ophthalmologically effective recommended for different formulation and delivery techniques The amounts are more fully described herein.   Introduction   Until the present invention, thyroid hormones have been used to synthesize hyaluronic acid ("HA") It is not recognized that can directly regulate intraocular pressure (“IOP”) won. Traditionally, the role of thyroid hormone in eye physiology has not been understood Was. Thyroid hormone in sufficient amount to exert the relevant physiological effect in the eye Whether it crosses the blood-brain barrier; ocular cells, such as trabecular meshwork cells, Whether you have the thyroid receptor required to specifically mediate the signal And thyroid hormone directly mediates hyaluronan secretion or synthesis by trabecular meshwork cells Or whether the IOP could be modulated.   Prior to the invention described herein, systemic levels of thyroid hormone Changes in the IOP due to change were assumed to be indirect or direct. Even so, changes in IOP induced by such thyroid hormones are Is thought to be mediated through adrenergic α or β receptors located within Had been. This time, the interpretation of the present invention is, for the first time, Recognize the important role of receptor. The thyroid receptors in the eye are It provides an opportunity to directly mediate the effects of intrathyroid thyroid hormones. In particular, Thyroid receptors in human trabecular meshwork ("HTM") cells are maintained by HTM cells It can potentially mediate the amount of extracellular HA, which, in turn, or) can affect the outflow.   Normally, IOP is maintained through aqueous humor secretion by the ciliary body and trabecular meshwork. Requires that all aqueous humor be balanced. The most important determinants of 10P One is the outflow of aqueous humor. A major part of the aqueous humor outflow is through the trabecular meshwork And is directed into the Canal of Schlemm, which passes through the collecting pipe. It is discharged into the episcleral vasculature tube of the episclera. Liquid The fluid traverses the trabecular meshwork before exiting the eye and entering Schlemm's canal. Pillar network That is, the network of secreted glycosaminoglycans ("GAGs") Hyaluronan and trabecular meshwork cells lining the meandering exit area extending into Schlemm's canal Consisting of The HTM cells produce or secrete GAGs, such as HA. HA is Solution hydraulic resistance increases the force required to transport liquids across distances HA can potentially be an important determinant of IOP, especially Acts as a rising IOP. Thus, one embodiment of the present invention reduces HA secretion or synthesis. By administering to the human trabecular meshwork cells an ophthalmologically effective amount of STH, It is designed to advantageously regulate HA secretion from trabecular meshwork cells in the eye. The present invention In one embodiment, for example, a method directed to glaucoma treatment does not necessarily inhibit HA secretion. No.   As a non-limiting introduction to the scope of the present invention, the present invention provides useful methods, compounds and combinations. Provides at least five categories of products:   1) For treatment of medical conditions related to thyroid hormone or GAG production or function A method directed to identifying a therapeutic compound of   2) Medical conditions related to GAG or hyaluronic acid, especially when symptoms occur in the eye A method directed to regulating GAG or hyaluronic acid production or activity in the condition,   3) a method directed to the treatment of glaucoma using STHs,   4) Compositions involved in delivering STHs to the eye, such as synthetic thyroid hormones Esters and diesters, and   5) Compositions related to the methods described in 1-3 and the compounds described in 4.   For example, one aspect of the invention is:   1) Screening of compounds using cells secreting GAGs or HA, for example, HTM cells Ning,   2) GAG or H in the presence of the test compound compared to the absence of the test compound A measurement of changes in production, and   3) Useful for treatment of medical conditions related to GAG or HA production, such as glaucoma Selection of appropriate compounds, Defines a method for identifying compounds useful as therapeutics, eg, therapeutic agents for the back of the eye.   Another embodiment of the invention provides for applying an effective amount of STH to a cell, such as a cell, for example. A method for regulating the production of GAGs by HTM cells is defined. STH applied is thyroid GAG production usually decreases when it is an agonist or partial agonist of the hormone Will be done. Another embodiment of the present invention minimizes the systemic effects of applied STH. Increases corneal passage after being applied topically to be metabolized in the eye STHs that can be directed to the eye using an ester derivative. above Methods, combinations of compounds and compositions are also contemplated. other Methods, compounds and compositions are also more fully described herein.   Drug discovery: screening for modulators of hyaluronic acid secretion or IOP Method   The present invention relates to thyroid hormone receptor-containing cells and cells associated with GAG production. The process involves compounds that are useful as therapeutic agents, e.g., compounds that modulate GAG production. Be a useful component of an in vitro or in vivo method for determining Understand. Typically, such identification methods allow for high-throughput automated screening Screening assay systems. Compounds useful as therapeutics The method for identifying a substance is as follows: 1) contact a cell with GAG-secreting cells, for example, trabecular meshwork And 2) while the compound has a concentration of less than 10 micromolar, Detecting binding of the compound to the cell. Detection of such bindings Are related functions (such as those described herein or developed in the art) For example, methods for assaying HA production) can be included.   HTM cells are preferred for screening for ophthalmologically active compounds However, other cell types can be readily used in place of HTM cells. Can be. In particular, such cells may be 1) endogenously invented or transferred genetically Thyroid receptor expressed from offspring, and 2) endogenously expressed or transferred Proteins involved in the production of GAGs, e.g., HA synthetase This is the case if it contains ze. For example, dermal fibroblasts can be used in such assays And would inhibit the overproduction of GAGs in the skin When screening for compounds or when HTM cells are obtained from a sample A particularly desirable choice as a convenient alternative to HTM cells when not possible You. In addition, non-HTM, GAG producing cells can be used in such screening assays. Alpha thyroid receptor subtype in the production of GAGs mimic any possible interaction between beta thyroid receptor subtypes Then, cells having the same thyroid receptor type as HTM cells are selected or combined. It is desirable to produce through replacement technology.   Human trabecular meshwork (HTM) cells are used to screen compounds, e.g., STHs If so, the HTM cells are used herein or in Polansky et al., Trabecular meshw ork cell culture in glaucoma research, Ophthalmology, 1984; 91: 580-595 ; Polansky et al., Human trabecular cells I: Establishment in tissue cul ture and growth characteristics, Invest. Ophthalmol. Vis. Sci. 1979; 18 : 1043-1049; Alvarado et al., Human trabecular cells II: Ultrastructural  characteristics of cultured trabecular cells, Invest. Ophthalmol. Vis. Sci., 23: 464-478 (1982); and Polansky et al., Studies on human trabecula. r cells propagated in vitro, Vision Res., 1981; 21: 155 (all of these May be prepared as described in US Pat. it can. For the screening assays described herein, third to fifth Using passage HTM culture from insulated stock, approximately 10,000 cells / cm^TwoIn plate And, for example, 10% fetal calf serum (FCS) to obtain a stable endothelial-like monolayer. Until confluence in Dulbecco's Modified Eagles (DME) medium containing 7-10 days (post-co nfluency). GAG, HA or protein synthesis, or transcription Saying can be performed, for example, using these stable HTM monolayer cultures. Trabecular meshwork cells are isolated from the eyes of rabbits, rats, mice, pigs, cats or monkeys. And can also be used for the assays described herein.   Assays of STHs for HTM cell division were performed using DME medium containing 10% FCS. It can also be evaluated during the cultivation of the nutrient. STH treatment converts 2,500 HTM cells /cm^TwoAt the day following the day of plating. The effect is logarithmic During the growth phase (7 days) and after the control culture has reached confluence (this is due to the HTM (Depending on the cell line and the serum in the culture medium, varies between 3-6 weeks) You.   The cells used in the above screening assays, e.g., HTM cells The binding of a compound, e.g., STH, can be detected using various methods. You. Direct detection of the amount of compound bound to cells, e.g., isotope, spectroscopy Use of binding assays with compounds with measurable or fluorometric labels Can be. Direct detection uses whole cells, isolated nuclei or chromatin or a combination thereof. To intracellular receptors in human trabecular meshwork cells, such as thyroid receptors Includes measurement of specific binding of the compound. Determination of thyroid hormone binding or analog binding The amount can be achieved using methods known in the art or Can be emitted. These are described in Schwartz, H .; L. et al., J. Amer. Biol. Chem. 1 7: 11794-11799 (1992), which is incorporated herein by reference. Including the methods used. The direct detection method is a synthetic thyroid hormone for its receptor. Affinity, esterified or non-esterified derivatives of the thyroid Useful in assessing the ability to bind to glandular receptors and assess STH uptake Would.   Alternatively, methods for indirectly detecting the amount of bound compound, such as receptors Gene constructs, DNA transcripts, RNA levels, proteolysis or synthesis, complex sugar degradation Or a functional assay to detect the effect of STH on synthesis and GAG synthesis or degradation. Can also be used . Other functional measurement methods described herein may also be used for functional assays. Can be used. Compound binding by both in vitro and in vivo methods Can be used to assay   For example, to identify compounds that modulate aqueous humor resistance within the trabecular meshwork of the eye, such as STHs Is desirable. In this case, in the presence and absence of the compound to be tested, By measuring changes in GAG production or hyaluronan secretion from reticulocytes Detecting the binding of a compound to trabecular meshwork cells, eg, HTM (human trabecular meshwork) cells Would be advantageous. Such controls may be used in any of the assays described herein. And other controls, such as functional thyroid receptors The effect of STH using or without added cells without detectable levels of By blocking, they can be easily interchanged to achieve specific detection. It will be appreciated that In addition, control values and control values Changes in the well-defined Will be understood in Say. In this case, it is necessary to perform a constant control. Will not. Because the control value and the change from that value have been established, And the experimental value can be appropriately measured in the absence of the control .   Once the compound, e.g., STH, is found to bind to cells, e.g., HTM cells If so, useful modulators of cell activity, such as GAG production, can be selected. Selection criteria are usually based on the degree of regulation produced by the tested compound. In the case of GAG production or HA secretion (or production), the compound usually blocks GAG cell production Will be selected based on their ability to reduce or reduce. Such a compound Substance is caused by inappropriate GAG production or HA secretion May be useful in the treatment of clinical symptoms. Such compounds may cause reduced intraocular It may also be useful in treating water spills or glaucoma. Typically for control GAG production By comparison, at least about 10%, preferably at least about 30%, and more preferably Compounds that suppress GAG production by at least about 70% are selected as useful compounds Will be. The above percentage inhibition criteria are based on the assays described herein. Other assays used in the assay, such as detecting HA secretion, detecting HA binding or measuring IOP Can also be applied.   More specific selection criteria identify compounds that more specifically modulate cellular processes Can be used to advantage. Tested for that receptor Affinity of a compound can often determine its specificity It will be understood that. Therefore, high apparent or real affinity It is desirable to select a compound that binds to the receptor or regulates its function. No. For STHs, the T for the thyroid receptor_Three(0.024nM under physiological conditions) Is T_Four(Affinity of 0.26 nM under physiological conditions) is preferred. New The thyroid gland for long-lasting and selective effects STHs Deduced from higher affinity or functional assays for the receptor Select STHs with apparent affinity, such as 0.1 nM or less under physiologic conditions It is desirable. To achieve the desired results as described above, a new compound or Known STHs can be synthesized and screened at a given concentration . Typically, the synthetic thyroid hormone is less than 1.0 micromolar, preferably 0.1 GAG production at concentrations below sub-micromolar, and most preferably below 0.01 micromolar Or it will reduce hyaluronic acid secretion. Especially if the STH has a higher ED50 Higher concentrations, for example 10-75 μM, can be used You. The percentage inhibition criteria discussed herein meet the criteria for concentration selection described above. Can also be used.   Identifying compounds useful for modulating cellular processes, such as GAG production, Both in vitro and in vivo screening assays can be included. Typically, compounds are first screened using an in vitro assay. And then screened using an in vivo assay. Imbi The bo assay and the measurements described herein may be useful for useful compounds, such as STHs. And can be used to further select. For example, in vitro Say also binds STH, which inhibits 40% of HA secretion with an apparent affinity of 10 nM. And STH localizes the synthetic thyroid hormone to the mammalian eye. It is further assayed by specific application. Alternatively, this in vivo update Say can be used alone to identify compounds.   Methods for treating medical conditions and glaucoma associated with GAG production and hyaluronic acid secretion   The present invention provides that thyroid hormone and STHs can be used to produce GAGs from cells, such as trabecular meshwork cells. Understand clearly that production or HA secretion can be directly regulated. Therefore The method according to the present invention provides for determining the amount or activity of a substance or cellular process in a desired cell or tissue. Modulating GAG production or HA secretion using compounds that reduce Pointed. Preferably, such cells are trabecular meshwork cells, ciliary body cells of the eye, ocular Endothelial cells, non-ocular endothelial cells, hair follicles and skin and internal organs Would be fibroblasts. In most cases, non-systemic application of STH is preferred. However STHs can also be applied systemically.   For example, the present invention relates to a method of inhibiting hyaluronic acid secretion, Synthetic thyroid formo inhibits hyaluronic acid secretion from cells that secrete luronate Non-systemic application of an effective amount of Of the above synthetic thyroid hormone after culturing at 37 ° C. for 3 days. Compared to human trabecular meshwork cells cultured in the absence of Mont, after culturing at 37 ° C for 3 days. Inhibits at least 10% of total hyaluronic acid secretion from cultured human trabecular meshwork cells, And the inhibition method. Preferably, the STH is located in the ocular tissue where the cells are isolated. At concentrations of 1-0.1 μM, whether in the trabecular meshwork or It will inhibit at least 10% of HA secretion by reticulocytes. Typically, this HA secretion inhibition method is: 1) T_ThreeSubconjunctival or intraocular injection, or 2) from natural tissue Isolated T_ThreeOr T_FourWill not include topical application of Non-systemic administration is local Includes both targeted and intraocular administration. Topical administration is, for example, a topical application. Non-surgically positioned holding device including rimmer and intraocular insert inserted under eyelid Sustained release from sustained release devices, topical application of ointments or creams, solutions, e.g. The intended cells, for example physiological saline solution and other solutions and substances suitable for the eye Or biologically and tissue-compatible with the tissue target, and Other extraocular delivery systems, including tablets, contacts, wafers or tablets including. Intraocular administration can be, for example, a surgically implantable or injectable intraocular Release device, which can be biodegradable or non-biodegradable. And those that can include a sustained release polymer. Applied non-systemically An effective amount and effective local concentration of the active compound and an ophthalmically compatible pharmaceutical composition. The ones that include the body are also described herein.   The present invention is not necessarily directed to inhibiting GAG production or HA secretion to create a therapeutic effect. It also includes aspects of the present invention that do not rely on it. Such as the present invention Embodiments include methods of treating glaucoma, methods of reducing or preventing elevated IOP, e.g., mineral col Elevated IOP associated with ticoid or steroid treatment, as well as tissue fluid flow or drainage For example, a method of increasing aqueous humor outflow in the eye. Preferably, these The method is therapeutically effective on a tissue or cell in need of treatment in a mammal, preferably a human. Relies on the non-systemic administration of large amounts of active compound.   For example, the present invention provides a method for treating an ophthalmologically effective amount of a synthetic thyroid hormone for the eye. Define a method of treating glaucoma, comprising administering to the required eye. Topical administration to the eye , Usually: 1) T_ThreeOr T_FourSubconjunctival or intraocular injection of 2) T isolated from natural tissue_Three Or T_FourTopical application or 3) T to the eye with cataract_ThreeOr T_FourAdministration, including Absent. Preferably, administration of STH to the eye is topical administration with an ophthalmically compatible pharmaceutical carrier. Or a polymer that provides sustained ocular sustained release of STH. Non-systemic administration using implants. Such controlled release of STH Can last from six months to one year. Such implants are osmotic pumps Can be a biodegradable matrix or sustained release device in the eye Wear. Implants allow the spread of synthetic thyroid hormone from contacts Also includes ocular contacts impregnated with synthetic thyroid hormone. For treatment of glaucoma The effective amount is usually 0.01 to 40 μg / eye / day, preferably 10 to 1,000 ng / eye. / Day, and more preferably 10-50 ng / eye / day. Preferably, topical application is Dispense at least one drop (eg, 30-50 μl) of a solution containing an ophthalmically compatible pharmaceutical carrier. Including giving. Other dosages and effective amounts and concentrations to request from the delivery system are: Are described in sufficient detail herein. Generally, 1-50 cells in the trabecular meshwork cells of the trabecular meshwork It is desirable to achieve a pM STH concentration. STH for thyroid receptors in the eye The apparent affinity of the When they are 10-100 nM (kd) and 100-1,000 nM (kd), respectively, 10-1,000 nM and 100-1 Higher concentrations of 0,000 nM are preferably achieved.   Typically, STHs administered to the eye (non-systemically or systemically) are usually In vitro or in vivo assays, such as the assays described herein Will be active. For glaucoma, synthetic thyroid hormone at concentrations of 10 micromolar or less Mon treatment is usually cultured in the absence of the applied synthetic thyroid hormone. Cultured trabecular meshwork cells at least 10% compared to hyaluronic acid secretion from isolated trabecular meshwork cells Will decrease the secretion of hyaluronic acid. Topical, if so desired The effect of locally applied STH can be measured before and after topically applying STH or as described herein. During the course of treatment and dosing appropriate for the individual's responsiveness to the treatment as described in It can be monitored by measuring the internal pressure. Serum levels of STHs It can also be measured to avoid unwanted systemic effects.   In the case of glaucoma treatment, this may be due to the activation or inactivation of systemically placed thyroid hormones. It is preferable to administer STHs non-systemically to avoid any systemic effects. Would be. The invention includes tissue treated with non-systemically applied STHs . For example, the present invention provides a composition comprising synthetic thyroid hormone in trabecular meshwork cells. Here, synthetic thyroid hormone does not cross the blood-eye barrier. Inside the cell and Is the location within the cell that is not in fluid contact with the extracellular environment, for example, the cytosol of the cell. , Nucleus or other organelle. The blood-eye barrier separates eye tissues from blood. An organization that is responsible for doing something. Preferably, STH in the corneal epithelium is synthetic thyroid It is an esterified derivative of a hormone. These cells can be from explants or Trabecular meshwork cells, ciliary body cells and endothelial cells of the eye in the living eye or any other mentioned herein Cells or tissues. Typically, the concentration of STHs in cells is low. Both would be 0.04 ng / dl. The amount (free and bound) and concentration of STH in cells Can be measured using methods known or developed in the future. This These methods are described in Oppenheimer, J. et al. H. et al., J. Amer. Clin. Invest., 75: 147-154 (1985); Schwartz, H .; L., et al., Endocrinology, 113: 1236 (1983) (this Both of which are incorporated herein by reference). Normal, The concentration of free STH in the cell is at least 1 pM, more preferably at least 10 pM And most preferably at least 100 pM.   Ophthalmic steroid treatment and use of trans-steroids are more frequent and Adverse side effects, such as elevated IOP. Interest in this side effect is a strong step. Restrictions on long-term ocular use of both loid and less active steroids ing. This is particularly the case for incurable blindness due to unrecognized rising IOPs. Many cases have been reported. The methods and compositions of the present invention provide an IOP free When a proper rise is measured or such a rise is expected, steroid therapy Can also be used to treat patients.   Elevated IOP can also occur in other medical conditions, e.g., after intraocular surgery. -It can also occur after treatment or trauma. IOP is the one described in this specification Can be regulated by STH prophylactically or after treatment or after trauma it can.   IOP (P in mmHg_o) Is directly proportional to the aqueous humor secretion rate (F in μl / min) Illustratively, and varies inversely with aqueous humor outflow capacity (C):           P_o= F / C + P_e Where P_e= Episcleral venous pressure (mmHg). P_eIs more specific for episcleral venous pressure P as another abbreviation_vTakes precedence over Administration of STH to the eye, especially non-systemic administration, The physician increases the aqueous humor outflow capacity (C), and the mammal or patient in need of the treatment Allow to decrease the rising IOP. Such treatment, as well as herein Other procedures described are, for example, related to onset of glaucoma, trauma, or surgery Can be used prophylactically to prevent increases in IOPs Wear. STHs may be applied as described herein for other treatments. it can.   Systemic treatment of glaucoma is also contemplated. Such systemic treatments are well known in the art. Blood T as is known_ThreeAnd T_FourEnsure that levels remain at non-toxic levels T for_ThreeAnd T_FourCan be merged with blood level monitoring and book Methods known in the art, for example, Murphy, B .; P. et al., J. Amer. Lab and Clin . Med., 66: 161-167 (1965). it can. In general, such treatment will reduce the medical symptoms of elevated IOP when the patient is identified. Have hypothyroidism-or thyroid if treatment is not required -Will not be used with hormone replacement therapy patients. Preferably elevated IOP Patients in need of STH treatment for thyroid gland will be hyperthyroid. Systemic by STHs Patients in need of treatment may also have_ThreeOr T_FourOr STHs thyroid It can withstand systemic levels of glandular hyperactivity. Instep administered systemically The daily dose of thyroid hormone is T_ThreeAbout 0.2 μg to 2.5 μg / kg / day (preferably , 0.2 μg to 1 μg / kg / day) and T_FourAbout 0.1 μg to 10 μg / kg / day ( Preferably, it will be in the range of 1 μg to 4 μg / kg / day). STHs are T_ThreeWhen T_FourWill vary in molecular weight from It may be appropriate to adjust the amount of STH performed. STHs ’ability to reduce IOP , In vitro and in vivo to establish a proper dosing regimen_ThreeAnd T_Four Can also be compared. At higher daily doses of STH, tapering or once daily It may be preferable to administer a daily dose of.   The treatments described herein may be associated with elevated IOP medical conditions, such as glaucoma or intraocular elevation. It can also be used to prevent blood pressure. Patients susceptible to elevated IOPs It can be effectively treated before the expected IOP rise occurs. Such a patient Persons include patients undergoing intraocular surgery or laser procedures.   Synthetic thyroid hormone   The present invention includes two classes of novel compounds. One class of compounds is the class of STHs. Esters of known synthetic thyroid hormones, including no- and di-ester derivatives Including conductors. This first class of compounds is based on the ST class known in the art. Does not include ester derivatives of Hs. However, such a compound is also used in the method according to the present invention. Can be used for The second class of compounds is the class of new STHs and their Including steal derivatives. Typically, the ester derivative has the formula:           X1-OC (O) -Y Wherein X1 is as defined below, and Y is a group compatible with the ester And Y is an alkyl, alkenyl or aryl group. Having from 2 to 20 carbon atoms. ｝ Medium, 1, 2, 3 or less STH with an ester group in the upper position. Typically, the Y group is 40 in size. Less than 30 atoms, and usually less than 30 atoms in size, and preferably It is hydrophobic and has no charge.   Di-ester derivatives are typically of the following formula (I):           X1-OC (O) -BC (O) -OX2 Wherein B is a linker (usually less than 6 carbon atoms in length); Where X1 and X2 are of the following formula (I): In the STHs ｛formula,   R₁Is CH_Three] CO_TwoH, -CH_TwoCH [NH-FMOC] CO_TwoH, -CH_TwoCH [NH-tBOC] CO_TwoH, -CH_TwoPO -CH_TwoCH [NHCO (CH_Two)_FifteenCH_Three] PO_ThreeH_Two, -CH_TwoCH [NH-FMOC] PO_ThreeH_Two, -CH_TwoCH [NH-tBOC] PO_ThreeH_Two , -CH_TwoSO_ThreeH_Two, -CH_TwoCH_TwoSO_ThreeH_Two, -CH_TwoCHNH_TwoSO_ThreeH_Two, -CH_Two FMOC] SO_ThreeH_Two, -CH_TwoCH [NH-tBOC] SO_ThreeH_TwoAnd   R_TwoIs   -H, halogen, CF_Three, OH, NH_Two, SH, CH_Three, -Et,   R_ThreeIs   -H, halogen, -CF_Three, -OH, -NH_Two, -N_Three, -SH, -CH_Three, -Et,   R_FiveIs   -H, halogen, -CF_Three, -OH, -NH_Two, -N_Three, -SH, -CH_Three, -Et,   R₆Is   -H, halogen, -CF_Three, -OH, -NH_Two, -SH, -CH_ThreeAnd   R '_TwoIs   -H, halogen, -CF_Three, -OH, -NH_Two, -N_Three, -SH, -CH_Three, -Et,   R '_ThreeIs   -H, halogen, -CF_Three, -SH, alkyl, aryl, 5- or 6-membered heterocyclic ring Aromatic or cyano,   R '_FourIs   -H, halogen, -CF_Three, -OH, -NH_Two, -SH, -CH_Three, -Et, or alkyl, aryl Or a 5- or 6-membered heterocyclic aromatic having a urea or carbamate bond Through O or N or S to R '_FourIs attached at the position,   R '_FiveIs   Alkyl, aryl, 5- or 6-membered heterocyclic aromatic, heteroalkyl, Reel, arylalkyl, heteroaryl, alkyl, polyaromatic, polyheteroaromatic And the above R ′_FiveCan be substituted by a polar or charged group,   R '₆Is   -H, halogen, -CF_Three, -OH, -NH_Two, -SH, -CH_Three, -Et,   X is   O, S, SO_Two, NH, NR₇, CH_Two, CHR₇, CR₇R₇Where R₇Is an alkyl, Aryl or a 5- or 6-membered heterocyclic aromatic,   And 'R₁-'R₆Group and R₁-R₆Any one of the radicals is replaced by a bond to B Can be replaced. ｝. ｝. Preferably, the GS Ter derivatives are two identical when hydrolyzed by intracellular hydrolases. Create molecules.   Preferably, the STHs have the following structure, Formula (II): Ｙ In the formula, Y is as defined above. ｝.   Usually, glaucoma patients or other types of patients requiring eye treatment are treated as eye drops. STHs are administered locally. However, when STHs are administered in the manner described above, Only about 0.1-1.0% of the dose is typically absorbed by the eye and A significant amount of the above dose can be transferred into the bloodstream. Low absorption of STHs in the eye The rate can be reduced by using at least three STH ester derivatives Can be caused by the following factors: 1) Ophthalmic solution flows immediately from the surface of the eye 2) STHs are rapidly absorbed into the bloodstream through the conjunctiva and / or nasal mucosa And 3) STHs poor transcorneal passage. STHs are absorbed into the eye through the cornea . In the cornea, STHs first absorb into the epithelial layer on the surface of the eye, which contains cell membrane lipids. Will be collected. STHs with charged groups have reduced lipid solubility, Diffusion across the skin can be reduced. The lipid solubility of STH (eg, S If improved (through tellurization), the passage of STHs through the corneal epithelium is enhanced and Provides greater delivery to woven cells. The cornea is the stromal and internal cornea Delivers STHs through the skin into the anterior chamber fluid and into trabecular meshwork cells and ciliary cells It can also act as a storage site. In addition, STH energy in cells, such as trabecular meshwork cells, The hydrolysis of stell derivatives leads to their non-hydrolyzed STH ester derivatives Create a constant diffusion gradient for Of intracellular non-hydrolyzed STH ester derivatives If the concentration is ester hydrolysis and pharmaceutical form Non-hydrolyzed STH ester derivatives are continuously maintained in cells Spread. On the other hand, STH in a hydrolyzed form accumulates in the cell. Obedience Thus, ester derivatives of STHs enhance the permeability of STHs and can be used in target tissues and cells. Will promote the accumulation of STHs.   Stability of STH ester derivatives should be tested in a buffer hydrolysis system Can be. The ionic strength (μ) of the buffer is usually 0.5. 4.0, 6.0, 7.4 and Used at a pH value of 9.0: using temperatures including: 37 ° C, 50 ° C, 60 ° C and 70 ° C Can be Half-life (T¹/_Two) Is tested in a buffer solution with a specific pH Calculated for the decomposition constant k obtained for each of the compounds Yes (T¹/_Two= 0.693 / k; k = 2.303 x kk, where kk is the remaining as a function of time Is the angle coefficient of a plot illustrating the logarithm of the present ester).   The storage stability of the ester derivatives of STHs under different storage conditions was To determine the decomposition constant, and calculate the decomposition temperature, and Equation (1): Where log k is given as a function of [1 / T]. According to プ ロ ッ ト above plot By calculating the decomposition constant k at the desired temperature from the equation be able to.   From the decomposition constant (k) obtained at the desired temperature, 10% of the drug Storage life time t indicating the time spent_Ten%(T_Ten%= 0.104 / k) You.   The lipophilicity of the STH ester derivative is determined by the partition coefficient for the compound at pH 7.40 ( It can be tested by measuring P). Measure the concentration of the compound to be tested in the aqueous buffer case by HPLC. It can also be performed in an octanol-aqueous buffer mixture by measuring the degree. Wear. However, the partition coefficient of a highly lipophilic compound depends on its retention time. (RP) Liquid chromatography (HPLC) (Beckmann 116 pump and 166 UV detector) Marathon autom. Sample feeder).   The half-life of the enzymatic hydrolysis of the novel STH ester derivative is 37 ° C at plasma / buffer pH7 .4--measured in the mixture (80% -20%) and as known in the art Can be Corneal permeability of STH ester derivative is in the delivery phase (epithelial side) Transfer of the compound from the corneal to the acceptor side (endothelium side) of the diffusion chamber Can be assessed by monitoring the movement. For example, rabbit eyes Of the cornea can be used. Collected from the acceptor side of the diffusion chamber Samples of the prodrug and the released drug as determined by HPLC Provide both concentrations. Therefore, the degradation rate of the prodrug in the cornea and the prodrug The corneal penetration of the lag can be measured.   Pharmaceutical composition   The present invention is directed to methods described herein and known in the art. It can also include pharmaceutical compositions that can be used for Typically, the composition The product may contain STH, especially an STH ester derivative and a pharmaceutically acceptable carrier. it can. Compositions are often described herein and are known in the art. Adapted to the particular delivery mode provided. ST for use in the present composition The exact type and amount of H will depend, for example, on the particular drug chosen, its dosage form, , Standard versus sustained release, the condition to which the medicament is administered, and the mammal to be treated Varies depending on the size and type of Will be.   The compounds of the present invention may be used as described herein for cellular and physiological In order to regulate the process, for example, to prevent an increase in IOP or to actually lower an elevated IOP You can be chosen to make it happen. Preferably, such compounds of the invention Is 1 × 10 in aqueous or treated tissue of the eye^-7Enough to provide effective concentrations below M Large amount, preferably about 1 × 10^-11M to about 1 × 10^-8The amount of M, and more preferably about 1 × 10^-11M to about 1 × 10^-9At elevated IOP when used in the amount of M Including STHs that provide reduction or prevention thereof.   For example, the present invention relates to a composition, preferably applied to the eye, comprising a synthetic thyroid gland. And those comprising Lumon and an ophthalmically compatible pharmaceutical carrier. Preferably, Such compositions may be formulated in buffered saline with or without liposomes. Including esterified derivatives of synthetic thyroid hormone that can be administered by combination . An ophthalmically compatible pharmaceutical carrier can also include a biodegradable synthetic polymer. . Thyroid hormones and STHs can be used for long-lasting eyes using biodegradable polymers. It can be delivered with internal release. Raw approved for human use Physically degradable microsphere compositions are polylactic acids: poly (lactic acid), poly (glycolic acid) ), And poly (milk-coglycol) acid. Additional biodegradable formulations Include, but are not limited to: poly (anhydride-co-imide), poly (lactic-glycolic acid), Triethyl-2-cyanoacrylate, polycaprolactone, polyhydroxybutyrate Valerate, polyorthoester, and polyethylene oxide / polybutylene -Contains tetraphthalate. Intraocular implant or sustained release thyroid hormone composition Injection of intraocular pressure while avoiding or reducing the need for topical preparations as much as possible Will provide long-term control (over months to years).   In another embodiment, the composition according to the invention comprises a synthetic thyroid hormone, preferably Is an ophthalmic contact device impregnated with an ester derivative of synthetic thyroid hormone ( For example, contact lenses). For example, dried STHs or their esters Derivatives are supplied as tablets and include contacts with prodrugs or medicaments. Melted in the presence of reusable or disposable contacts for immersion Can be. After a suitable incubation time, e.g. Is rinsed from the incubation solution, and STH is Applied to the eye to allow diffusion into the eye. Ashton, P .; et al., J. Amer. of O cc. Pharm. 10: 691-701 (1994) Intraocular sustained release devices, including those described by It would also be advantageous to use chairs.   In general, formulations suitable for topical and intraocular administration will be prepared by techniques known to those skilled in the art. And can be administered in accordance with Distribution that can be oxidized The compound is preferably made anoxic by making all formulations under an inert gas. Prepared in the background. Finished formulations preferably protect them from exposure Storage in an opaque or brown container and in an inert atmosphere.   Aqueous polymer solutions, suspensions, ointments, and gels are preferably formulated topically Used for things. Aqueous formulation creates reservoir for dissolved therapeutic agent For this purpose, liposomes can be included. Particularly preferred among topical formulations are: A gel that enhances precorneal retention without the ocular inconveniences and impairments associated with ointments is there.   Topical ophthalmic or other topical formulations generally will contain from 0.001 to 1 in a suitable polymeric carrier. 0% by weight, preferably 0.05-1% by weight, and most preferably 0.05-0.6% by weight Should be included. Other preferred formulations have 0.001-0.009% by weight of therapeutic Agent. By those skilled in the art As can be appreciated, the amount of STH needed to reduce IOP or glaucoma Drugs or prodrugs that are less absorbed may cause a noticeable systemic effect. It is an amount that will not occur.   Suitable polymeric carriers include lightly crosslinked carboxy-containing polymers (eg, poly). Licarbophil, dextran, cellulose derivatives, polyether Includes Tylene glycol 400 and other polymeric demulcents.   Preferred systems are well known in the art, such as low cross-linked acrylic acid or the like. Including rimers. In a preferred embodiment, such a polymer comprises At least about 90% by weight, and preferably about 95% by weight, based on the total weight of the % To about 99.9% by weight of one or more carbonyl-containing monoethylenically unsaturated monomers. Prepared from other unsaturated, polymerizable carboxyl-containing monomers, e.g. For example, methacrylic acid, ethacrylic acid, β-methylacrylic acid (crotonic acid), cis -Α-methyl crotonic acid (angelic acid), trans-α-methyl crotonic acid ( Tigulinic acid), α-butyl crotonic acid, α-phenylacrylic acid, α-benzyl Acrylic acid, α-cyclohexylacrylic acid, β-phenylacrylic acid (cinnamic acid ), Coumaric acid (o-hydroxycinnamic acid), embellic acid (p- (Hydroxycoumaric acid) and others are used in addition to or instead of acrylic acid Can be done.   Such polymers have a low percentage based on the total weight of the monomers present. Page, i.e., from about 0.01% to about 5%, and preferably from about 0.1% to about 2%. Crosslinked by using a functional crosslinking agent. Some of the above crosslinking agents include non-polyamide Renyl polyether bifunctional crosslinkable monomers such as divinyl glycol; 2,3-dihydroxyhexa-1,5-diene; 2,5-dimethyl-1,5-he Xadione; divinylbenzene; N, N-diallylacrylamide; N, N-di Allyl methacrylamide and others are included. Also, two or more alkenyls per molecule Ether group, preferably two or more alkenyl ether groups per molecule, preferably Is the terminal H_TwoPolyalkenyl poly containing alkenyl ether group containing C = C <group An ether crosslinker comprising at least 4 carbon atoms and at least 3 hydroxyl groups Can be converted to alkenyl, such as allyl bromide, etc. Prepared by tellurization, for example, polyallyl sucrose, poly Allyl pentaerythritol and others are also included; for example, US Pat. No. 2,798,05. See issue 3. Diolefin non-hydrophilic macro with a molecular weight of about 400 to about 8,000 Mer crosslinkers such as insoluble di- and polyacrylates and diols and Methacrylate, diisocyanate-hydroxyalkyl acrylate Or methacrylate reaction product, and polyester diol, polyether Isocyanate-terminated pres derived from diols or polysilaxane diols Reaction products of polymers and hydroxyalkyl methacrylates, etc. Agents, for example; Mueller et al., US Pat. No. 4,192, See 827 and 4,136,250. Low crosslinked polymers, of course, Carboxyl as the sole monoethylenically unsaturated monomer present with the crosslinker Can be made from the monomer (s) containing the monomer. They are 40% by weight The following, and preferably from about 0% to about 20%, by weight of the carboxyl-containing monoester The thylene unsaturated monomer (s) is an acrylic acid ester and methacrylic acid Esters such as methyl methacrylate, ethyl acrylate, butyl Acrylate, 2-ethylhexyl acrylate, octyl methacrylate, 2- Hydroxymethyl-methacrylate , 3-hydroxypropyl acrylate, others, vinyl acetate, N-vinyl pyro Contains only physiologically and ophthalmologically harmless substituents, including ridone and others Replaced by one or more non-carboxyl containing monoethylenically unsaturated monomers Such an additional monoethylenically unsaturated polymer. For a broader listing of sum monomers see Mueller et al., US Patent No. 4,548,990. checking ... Particularly preferred polymers are low crosslinked acrylic acid polymers, The crosslinking monomer is 2,3-dihydroxyhexa-1,5-diene or 2, 3-dimethylhexa-1,5-diene.   The low crosslinked polymer used in the practice of the present invention is preferably, for example, It ranges in size from about 1 to about 30 μm, and preferably to the equivalent spherical diameter. Conventional to provide dry polymer particles ranging from about 3 to about 20 μm in Of free-radical polymerization catalyst of about 50 μm or less in equivalent sphere diameter It is prepared by suspending or emulsion polymerizing the above monomers to a dry particle size. Generally, such polymers have molecular weights estimated to be greater than 2,000,000. Will be distributed.   Where the dry particle size is considerably larger than the equivalent spherical diameter of about 50μm Formulated according to the present invention comprising polymer particles prepared by suspension or emulsion polymerization of The resulting aqueous suspension, when administered to the eye, has an equivalent spherical diameter of less than about 50 μm on average. Is less comfortable than a suspension in a composition comprising polymer particles where Or the same. Dry particle size much larger than about 50 μm in equivalent sphere diameter And then, for example, by mechanical grinding or grinding Reduced in size to a dry particle size of about 50 μm or less in equivalent sphere diameter Low cross-linked acrylic acid, etc. Limers do not perform as well as polymers made from aqueous suspensions. Exists Difference of such mechanically milled polymer particles as the only particulate polymer One possible explanation for the difference is probably to remove non-bridging branches from the polymer chains, Produce particles with sharp edges or protrusions or have a satisfactory delivery system By creating a particle size range that is usually too wide to give The spatial geometry or configuration of lowly crosslinked polymer particles larger than μm The crushing will destroy it. The wide distribution of particle size is related to viscosity-gelation Will hurt. In any case, the particles thus mechanically reduced are suspended Or more volume in aqueous suspension than particles prepared in an appropriate size by emulsion polymerization. It cannot be easily hydrated and also has a sufficient intraocular effect under the influence of tears Aqueous suspension according to the invention once gelled Less comfortable than gels produced intraocularly using However, there is About 40% by weight or less, for example about 0% to 20% by weight, based on the highly crosslinked particles % Of the above ground polymer particles, when practicing the present invention, has a particle size of about 50 μm or less. Sharpening dry particle diameter, can be mixed with solution or emulsion weight polymer particles . Such a mixture is particularly useful when the polymer particles thus milled are in a dry form. At an average value of about 0.01 to about 30 μm, and preferably at an equivalent spherical diameter. When having an average value of about 1 to about 10 μm, ophthalmic drug delivery is easier and more comfortable. A satisfactory viscosity level in the system, as well as a satisfactory sustained release of the drug into the eye Will also provide exits.   In another aspect of the invention, the particles have a narrow particle size distribution. Monodisperse particles Is the maximum viscosity of the ophthalmic drug delivery system for a given particle size And will give increased eye retention time U. Monodisperse particles having a particle size of 30 μm or less are most preferred. Good particle packing Is aided by a narrow particle size distribution.   These particles not only adhere to the above size limit, but also have a narrow particle size distribution. Obey. The above use of monodispersion of particles to help good particle packing, Creates a maximum increased viscosity during contact with the suspension and increases intraocular residence time You. At least about 80%, more preferably at least about 90%, and most preferably Means that at least about 95% of the particles are within the major particle size distribution of the band Should be, and as a whole (ie, inside and outside such a band Less than about 20%, preferably less than about 10%, and Also preferably, no more than about 5% of fines (ie, particles of size less than 1 μm) Child) should exist.   The average particle size is, for example, 6 μm from the upper limit of 50 μm, more preferably 30 μm. When reduced to a low size, the band of the primary particle distribution is also narrow, for example to 5 μm. It is also preferred that Preferred size for particles in the band of primary particle distribution Is less than about 30 μm, more preferably less than about 20 μm, and most preferably from about 1 μm to It is about 5 μm.   The aqueous suspension according to the invention is preferably based on the total weight of the aqueous suspension. From about 0.1% to about 6.5%, and preferably from about 0.5% to about 4.5% by weight. It can include a significant amount of surrounding low crosslinked polymer particles. It Are preferably physiologically pure, sterile water, preferably deionized or distilled. Or using ingredients that are not ophthalmologically harmful, and Physiologically and ophthalmologically acceptable pH-adjusting acids, bases or buffers, such as acids For example, acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, hydrochloric acid, and other bases such as Sodium hydroxide, sodium phosphate , Sodium borate, sodium citrate, sodium acetate, sodium lactate , THAM (trishydroxymethylamino-methane), others, and salts and Fur, such as citrate / dextrose, sodium bicarbonate, ammonium chloride Prepared from about 7.0 to about 7.4 neutral using a mixture of sodium and a mixture of the above acids and bases. There will be. However, the eye will tolerate pHs outside the neutral range, and Acidic or basic pH can also be used to facilitate drug solubility.   Aqueous suspensions are filled into preservative-free, single-dose, non-reclosable containers. Can be This means that a single dose of the drug is delivered to the eye one drop at a time The container is discarded after use. Such containers are, in particular, mercury Preservation of the corneal epithelium, as has been observed to result from ophthalmic medications containing preservatives Eliminates charge-related stimuli and sensitization.   Multi-dose containers can be used if desired. In particular, the aqueous suspensions of the invention The relatively low viscosity of the eye, as many times daily as needed, provides a precise dose This is because it allows the eyes to be seen. Such as preservatives must be included In suspensions, the preferred preservatives are chlorobutanol, polyquat salt Benzalium ruconium bromide, cetyl bromide and others.   Additives desirably included in topical formulations include sodium chloride, EDTA (edetate) Disodium), surfactants and preservatives such as (benzalkonium chloride) Including. The administration of the formulation to the eye will typically depend on the particular problem being treated, Will be performed 1-4 times per day.   Formulations for ophthalmic injection fall into two classes. For subconjunctival injection, one Generally, the formulation will comprise from 0.0001 to 1% by weight, preferably It should contain 0.001-0.1% by weight of the therapeutic agent. Any suitable carrier, preferably Can be a polymer carrier, such as dextran or polysorbate 80. it can. Other additives that may be desirably included in the formulation include edet 2 Thorium, sodium bisulfite and sodium sulfite. This formulation contains phosphate buffered saline, citrate buffered saline Saline, chondroitin sulfate, or a polymer carrier, such as sodium hyaluronate (Or hyaluronic acid), including purified polyacrylamide or polysorbate 80 Should. Other additives desirably included in ophthalmic formulations are sodium chloride Sodium hydroxide and hydrochloric acid. Used for Typically, 0.001-1% by weight, especially when in solution, is preferred. Or 0.01 to 1.0% by weight of the above agent.   When the active compound or prodrug is substantially in solution, its therapeutic action is Is immediately possible, and therefore lower concentrations are Can be administered to achieve an effective level without causing Activity When the compound or prodrug is substantially in suspension, the higher concentration is again Administered to achieve sustained effective levels without causing tissue intolerance Can be Therefore, in solutions, lower concentrations can reduce local tissue damage. Used to evade. In suspensions, higher concentrations are used. why If so, a lower dissolved amount is introduced for immediate activity.   To obtain the desired effective concentration for treatment for a particular prescription of a particular patient Dosage and effective amounts will be according to the desired clinical endpoint, and dosing regimen will be adjusted as appropriate. By knotting, it can be easily obtained. For example, non-systemic administration to the eye In the case of STHs Is less sensitive to systemic effects. Therefore, investment per day Adjustments in dosage and dosage per application interfere with the bioavailability of the drug Can be changed without low effects from organs such as the liver, intestines and kidneys Wear.   Clinical endpoints are easily monitored and compared to clinical subjects from comparable normal subjects. At the clinical endpoint or at the clinical endpoint measured prior to the procedure. And as known in the art. For example, positive In the normal eye, changes in aqueous humor secretion are caused by diurnal fluctuations. ), Aging, endocrine disruption, hydration, medicine, changes in intraocular pressure, such as surgical results Can be related to factors. However, such changes in aqueous humor secretion are Maintain a relatively constant intraocular pressure and measure only the IOP over the course of the procedure. As a small compensatory adjustment in the ease of spillage to give a slight effect on Always appear. As described herein, a normal IOP value indicates that the eye has an appropriate IOP. Elevated IOP to ensure reduction or reduction of STH administration or STH treatment when returning to OP level It can be used as a reference point to indicate a value. For glaucoma, the IOP is In the case of raised and open-angle glaucoma, ease of spillage is reduced. This is the IOP With greater fluctuations in intraocular pressure due to elevations in blood pressure and changes in aqueous humor secretion Sir. The IOP applies a force to its eyes that knurls or flattens it. Can be measured by Clinicians need to measure changes in the IOP The course of STH treatment can be better assessed.   IOP can be measured using an applanation tonometer. The preferred pressure is the cornea Measuring the force required to flatten the standard area (3.06 mm diameter) It is measured in the eye before and after the procedure using a dynamometer (tonometer). Applanation tonometer Do not replace a large amount of liquid ( About 0.5 μl) or does not significantly increase intraocular pressure. Independent of eye hardness. Typically, the eyes are 0.5% proparacaine (Ophthaine) , 0.4% Benoxynate (Dorsacaine), or a similar local anesthetic (tetracaine (Except for Pontocaine), the tears of which are isotonic sodium chloride Sterile fluorescein paper strips moistened with urea solution or distilled water To make it fluorescent. The IOP is then measured using a tonometer. Subject is horizontal Or it can be vertical. Various other tonometers and methods measure IOP Can also be used forExample   Example 1-Materials and Methods   The following materials and methods achieve the results obtained in the examples described herein. Can be used to Other materials and materials known in the art And methods can be used to achieve similar results. Cell culture   Primary, cultured human trabecular meshwork ("HTM") cells at passage 3-5 were used for all experiments. used. Cells are grown in 11% CO_TwoAnd 15% FCS (fetal calf serum) at 37 ° C, Genta Mycin, glutamine (2 mm), fanzizone (2.5 μl / ml), penicillin / s DME-H 16 supplemented with treptomycin (UCSF Cell Culture Facility) 1g / Cultured in L glucose (standard medium). 10% explants for 1-2 weeks without disturbing CO_TwoIncubated in a humidified incubator. By week 3 in culture The cells naturally migrated from the tissue. At this point, the medium was changed every other day, and Fibroblast growth factor (1 ng / ml) was added after each medium change. Cell mass is 10 Reached the total number of 00-2000 At times, the explants are removed and cells are STV (0.1% trypsin, 0.02% EDTA ) And trypsinized and plated on 60 mm dishes. The medium is collected Replace every other day until dense, usually 7 to 8 days after starting plating did. Confluent cultures were grown at 1 × 10 5 per sample in liquid nitrogen for future use.^-6 Frozen in cells or 1:32 split for experimental use or continuous culture The cells were subcultured at different ratios. HTM cells maintain morphology for more than 20 passages. Only However, in most experiments, HTM cells from passages 3-5 were used . These cells are substantially pure and can be made by methods known in the art, For example, Alvarado, et al., Invest. Ophthalmol. Vis. Sci., 82: 464-478 (1982) Can be prepared. Preferably, the cell has an effect on cell growth. Cultured to confluence before use in experiments to reduce. Non-confluent cells are typical Typically less than 80% congestion.   Ten^-2M-conserved 3,3'-5-triiodo-L-thyronine (T_Three) (Sigma, St. Lo) uis. MO) was prepared in 10% DMSO / 90% EtoH and stored at -20 ° C. Co valink-NH microwell plate (Nuna, Naperville, IL), N-hydroxy Sisulfone succinimide (S-NHS) (Pierce, Rockford, IL), hyaluronic Acid (INC, Costa Mesa, CA), 1-ethyl-3- (3-dimethylamino-propyl) ) Carbodiimide (EDC) (Sigma), O-phenylenediamine (OPD) (Calbiochem , San Diego, CA), Vectastain standard ABC kit (Vector, Burlingame, CA) Used for hyaluronidase and HA ELISA-like assays. Assay for hyaluronic acid   HTM cells were supplemented with 2 ml of 10% fetal calf serum medium without thyroid hormone. DME-H 16 in 1 g / L glucose and 10^-7M   T_Three6-well or 12-well (Corning, Corn) with or without the addition of ing, NY). Change the medium daily or as indicated, and Collected. The supernatant is concentrated and then the biotinylated hyaluronic acid binding protein 3 for hyaluronic acid ("HA") by ELISA-like assay using quality HABP Assays were performed in duplicate. Stern & Stern, Matrix, 12 (5): described in 397-403 (1992) Assays such as those described can be used. Assay for hyaluronidase activity   HTM cells were allowed to undergo insulin-transferrin-se over the number of days indicated in the figure legend. In 10 ml serum-free DME-H16 supplemented with rhenium, 10^-7MT_ThreeWith the addition of Without incubation, the cells were cultured in 100 mm dishes (Fisher, Pittsburgh, PA). Serum-free medium, Hyaluronidase inhibitory proteins normally present in serum and interference with hyaluronidase Used to prevent harm. Concentrate supernatant in centricon (Amicon) And ELISA using directly biotinylated HA (Stern &  Stern, Matrix, 12 (5): 397-403 (1992), the method of which is incorporated herein by reference. The incorporation) modification results in a hair in pH 3.7 citrate-phosphate buffer. Assays for luronidase activity were performed in triplicate. Assemble all samples in triplicate I did.   Example 2-Synthetic thyroid hormone, eg, T_ThreeInhibits HA secretion from cells   Find out if HA secretion can be regulated by synthetic thyroid hormone HTM (“HTM”) cells_ThreeWas cultured in the presence and absence of E. coli. T_ThreeSupply culture HTM cells cultured in the ground_ThreeCompared to cells cultured in media with Showed a gradual decrease in net hyaluronic acid secretion. See FIG. "Stripped" media is described in Samuel et al. (Samuel HH, Stanley , Cananova J: Endocrinology, 1979 Jul, 105 (1): 80-5) T using such an anion exchange resin method_ThreeRefers to the medium from which is removed. Scripture Formally, T in the removal medium_ThreeIs less than 0.1 pM. T_ThreeSupplemental and control media The difference in secreted HA concentration in the (confluent cells)_Three(Most Final medium concentration 10^-7M) could be observed on day 3 after exposure. Day 5 By time, compared to the supernatant from the control culture,_ThreeSecreted H in the supernatant from the supplemented culture There was a 2.95-fold decrease in A concentration. Differences in HA levels were observed by day 10 of culture. (The last day of the experiment). T_ThreeUntreated (less than 75% confluent) cells treated , Congestion T_ThreeIn medium compared to treated cells (each well contained approximately 250,000 cells) Produced even greater reductions in HA concentration. T_ThreeFor reduction of HA concentration by The half-maximal concentration was about 5 nM. For confluent cells, the dose response in FIG. See response curve.   Example 3-T_ThreeIncreases the activity of secreted hyaluronidase   T_ThreeIncreased secretion of hyaluronan in the presence of HTM cells to determine if they can also be partially regulated. The enzymatic activity of the secreted hyaluronidase is determined by T_ThreeMeasured in the presence and absence of . HTM cells are treated with serum hyaluronidase and / or hyaluronidase inhibition Serum-free for hyaluronidase activity experiments to eliminate confounding of sex proteins Cultured in medium. Secreted hyaluronidase activity is T_ThreeFor controls lacking additive T in comparison_ThreeIncreased in the supernatant from the supplemented unconfluent culture. By day 10, serum-free strips Despite the down regulation of HA production under the_ThreeSecretion in supplemented cultures There was a relatively 1.9-fold increase in hyaluronidase activity.   Example 4-CD44 isoform capable of modulating secreted HA levels Is present on HTM cells   To determine whether the CD44 receptor is expressed by HTM cells, Probe the D44 isoforms for CD44s, CD44E, CD44-v1 and CD44-v3 Can be measured using RT-PCR analysis using All isoforms are Splice alterations arising within nine external axons with different splice sites It is a variant. All four isoforms are expressed from HTM cells. RT-PCR results For experiments, cells (1-3 × 10⁶) Is harvested by scraping, Pellet, freeze immediately in liquid nitrogen, and until RNA and cDNA are made saved. RNA was used using RNA Track ™ (Biotecx, Friendswood, Tx) . This RNA was transferred to I000 U of Moloney Murine Leukemia Vir in a total volume of 60 μl. us Reverse Transcriptase (BRL, Gaithersburg, MD), 50 mM random hexagon Mer primer, 50 mM Tris-HCl (pH 8.3), 75 mM KCl, 10 mM dithiotrate , 3mM MgCl_TwoIn a reaction mixture containing 0.5 mM each of dGTP, dATP, dTTP and dCTP. Used as template for first strand synthesis of DNA. cDNA reaction at 37 ° C for 90 minutes And then diluted 1: 3 or 1: 7 in water pretreated with DEPC. Oh Ligonucleotide primers were diluted to 40 μM in water.   For RT-PCR, a modified hot start method was used. 3-5 μl of cDNA The volume was adjusted to include 0.8 μmol of each primer, 17 mM Tris-HCl (pH 8.3) and 80 mM KCl. Was added to 30 μl of the master mix. Add water to the cDNA reaction to a final volume of 40 μl Was added. All reactions were overlaid with light mineral oil and heated to 99 ° C. 10 minutes later The reaction was cooled to 94 ° C and 7 mM MgCl_Two, 1 mM dATP, dTTP, dGTP And dCTP, and 1.2 U of Taq Polymerase (Perkin-Elmer Cetus, Branchburg, NJ) ) Is added directly through the upper layer of the above oil. Added. The cycle parameters were: 95 ° C, 60 seconds, 55-57 ° C, 30 seconds, 72 ° C, A final extension of 15 minutes at 75 ° C. for 30 seconds. Reactions are automatically programmed 32 to 34 times using a thermocycler (Perkin-Elmer Cetus, Norwalk, CT) I returned. The RT-PCR product was converted to 2% Low EEO agarose (Fisher Scientific, Pitt burgh, PA). Run ethidium bromide (EtBr) gel at 200V for 45 minutes , Visualized with UV light, and photographed.   Oligonucleotides for RT-PCR can be used to efficiently increase all of the CD44 isoforms. Made to exons within the CD44 gene in a manner that would allow for width. Oligo Nucleotides as forward [F] or backward [R] and where they are directed Named by Roxon Exon:   Such a CD44 isoform binds to HA and, at least in part, resects secreted HA. Regulates sceptor-mediated endocytosis. In this pathway, HA Internalized via the sceptor and lysed for degradation by hyaluronidase Directed into the sosome. Furthermore, CD44s and CD44E are independent of endocytosis It has been shown to anchor HA and other GAGs to the cell surface.   Example 5-T_ThreeReduces expression of the CD44 isoform   T_ThreeIs secreted HA turnover by HTM through other splicing of CD44 receptor gene CD44 expression is described herein to determine whether it affects conversion. Monitor using RT-PCR Can be HTM cells replenished for 18 hours_ThreeCD44E when exposed to And there is a dramatic decrease in the expression of the CD44-v3 isoform. Therefore, CD44 expression T_ThreeInduced reduction is inconsistent with reduced binding of HA and other GAGs to trabecular cells Absent.   Example 6-Human eye is aqueous T_Fourincluding   Human aqueous humor is T_FourTo determine if it contains_FourA known thyroid disease Dialysis technique from aqueous humor obtained during cataract surgery for patients without dialysis ( Corning Nichols Institute, San Juan Capistrano, CA). The pooled sample of aqueous humor sample had a free T of 0.4 ng / dL._FourLevel indicated. T_Three Was not measured at all in the same sample.   Example 7-Human TM cells of the eye have thyroid hormone receptors   Determine if human eyes, especially HTM cells, contain thyroid hormone receptors Thyroid hormone receptor and thyroid receptor using HTM cells -Thyroid hormone levels using RT-PCR analysis or immuno-histochemistry for nucleic acids It was measured using an antibody against the scepter. Thyroid hormone receptor α1, 2 and β1 were seen using both assays and nonspecific binding was relatively low. Was.   Example 8-T in vivo in the eye_ThreeTopical administration into the cornea and posterior chamber T_ThreeBring about the spread of   To determine the ability of synthetic thyroid hormone to cross eye tissues, I¹²⁵-T_Three Was administered to rabbit eyes in vivo and radioactivity was measured in aqueous humor. Simple Simply put, the following procedure is used to measure thyroid hormone passage into the eye 1) Anesthetize rabbits with 2: 1 ketamine: xylazine, 2) 1 Apply a drop of local anesthetic to the eye and 3) use the Balanced Salin Solution (Alcon) Rinse and dab dry 4) I in 100% EtoH solution¹²⁵-T_Three I in 30% DMSO in 30 μl PBS (phosphate buffered saline)¹²⁵  T_Three 5) 30 minutes to 1 hour for alcohol solution and DMSO solution T for 45 minutes for liquid_ThreeAbsorb solution and 6) T for about 30 seconds in control eyes_Three Absorb the solution and 7) remove 60 ml of aqueous humor with a disposable syringe before removing 50 μl of aqueous humor Rinse eyes with sterile water and 8) release from aqueous humor sample in gamma counter Measure the shooting power. Autoradiography with local I¹²⁵-T_Three2 hours of administration of This was performed on rabbit eyes that were later excised. Label is the aqueous humor outflow channel of the eye Had been concentrated. This is due to the locally administered T_ThreeCan be connected to the trabecular meshwork And that it can be done. Using such a method, the applied T_ThreeAbout 0.125% of the water diffuses into the aqueous humor after 45-60 minutes, and the diffusion Is DMSO T_ThreeIndependently of whether it is used as a solvent for I have. Therefore, T_ThreeCan diffuse across the corneal diffusion barrier, and Therefore, access to the aqueous humor and trabecular meshwork is possible.   Example 9-T in vivo in the eye_ThreeTopical administration reduces intraocular pressure   Synthetic thyroid hormone for intraocular pressure ("IOP"), such as T_ThreeTo see the effect of In order, T_ThreeWas administered topically to rabbit eyes in vivo. Normal pigmented Dutch The heron was used for the IOP experiment and T_ThreeWith PBS containing 30% DMSO medium four times a day 1mM T_ThreeThe solution was topically administered to the right eye of a rabbit as 30 μl of a sterile solution. 30% DMSO medium The body was applied to the control right eye four times per day. At predetermined time, IOP pulmonary incision technique Measured using In clinical situations, lower T_ThreeThe amount of Systemic toxicity or disturbance, such as tachycardia (tachyca rdia) will be administered to reduce the effect.   5 rabbits have at least some detectable low in IOP within 3 weeks of dosing Shows below, and one rabbit has T_Three3 weeks after administration, equivalent to a 25% decrease in 10P Decreased. The decrease observed at 10P in both eyes was not a T given_ThreeCan be related to systemic absorption of Measured T_ThreeSerum Levels increased in three rabbits two weeks after topical treatment (88-160 ng / dL). With a normal level of 176-427 ng / dL). 1 rabbit, probably topical High serum T resulting from administration_ThreeT from concentration_ThreeDied due to toxicity. T_ThreeThe toxicity of As described herein, use of low activity STHs or low concentrations of STHs Amount of synthetic thyroid hormone applied using prodrug ester derivatives Can be avoided by reducing.   Example 10-Hydraulic conductivity of cultured human trabecular meshwork cells: in vivo outflow of aqueous humor B model   As a measure of the ability of thyroid hormone to perform HTM in vivo, Is T_ThreeTraverses a monolayer of HTM cells in vitro Assess if it can work to cut and increase liquid flow directly did. We used an in vitro model to measure hydraulic conductivity . This model measures the hydraulic conductivity of a confluent monolayer of HTM cells.   Cultivate a monolayer of HTM cells on a Millipore filter in a polystyrene holder. Nourished. The pressure that develops when a liquid flows across a known resistance through this holder Place in the device to measure the difference. Two pressure gauges, a known resistor 1 upstream, and downstream of its known register, but containing HTM cells The one immediately upstream of the filter holder measures the pressure difference. This filter Is at atmospheric pressure, so the pressure in the second gauge is the perfusion pressure of the cells. The hydraulic conductivity is Q / PA ／, where Q is the flow, P is the perfusion pressure, and A is the monolayer surface Area. }be equivalent to.   Confluent HTM cells were treated with 2 mM glutamine, 1 nM penicillin, and 1 nM streptomycin. 1 cm Millipore filters in DMEM-H16 containing Syn (Millipore Corp., Bedf ord, MA) for 4 or 8 days. In addition, this medium contains activated carbon and mixed clicks. With on / anion exchange resin (AG 501-X8 Resin, Bio-Rad, Hercules, CA) From 10% fetal calf serum that has been hormone-depleted by incubation Was made up. 10 confluent HTM cells^-7MT_ThreeCultured for 4 or 8 days with or without . Control cells were treated with an appropriate amount of ethanol vehicle. Hydraulic transmission of each filter Conductivity was achieved by perfusing the cells with the same medium in which they were cultured for 30 minutes. Measured over time and μl / min / mmHg / cm^TwoWould be represented as Up All of the experiments were performed in quadruplicate.   After 4 days of treatment, T_ThreeHTM cells incubated with_ThreeNon-existence of 1.5 to 2 times more water than HTM cells cultured underneath It had mechanical conductivity (FIG. 3). However, this difference is statistically significant There was no (Wilcoxon test). After 8 days of treatment, T_ThreeIncubate with HTM cells_Three2-3 times more statistically significant than HTM cultured in the absence of It had large hydraulic conductivity (p = 0.0286, Wilcoxon test) (FIG. 4). Therefore, T_ThreeActs directly on the confluent layer of HTM cells, preventing fluid flow. It reduces their resistance and increases their hydraulic conductivity. further, The magnitude of this effect increases epinephrine, the outflow of aqueous humor, and Similar to previously reported studies with anti-glaucoma agents that reduce intraocular pressure . These results are significant. Because in vitro measurements of hydraulic conductivity Ophthalmic medications for aqueous humor outflow in vivo, such as epinephrine and corti This is because it correlates well with the observed effects of costeroids. Therefore, T_ThreeIs in It increases the hydraulic conductivity in vitro, so it can be used in patients with glaucoma. Can increase aqueous humor outflow and decrease intraocular pressure.   Example 11-T_ThreeIs the extracellular matrix of cultured human trabecular meshwork cells. Regulate adhesion   We are T_ThreeHTM cells cultured in the presence of_ThreeHTM cultured in the absence of It has been shown to produce less hyaluronic acid than cells. Hyaluronic Acid (HA) usually binds to the cell surface receptor, CD44, Interacts with cells by forming T_ThreeHTM cells bind to HA and To assess the ability to assemble extracellular matrices We see that HTM cells grown in culture bind to HA and see it The ability to assemble into an extracellular matrix was evaluated. This assay system From the area surrounding the cell By taking advantage of its ability to eliminate added formalin-added red blood cells, Visualize the extracellular matrix of the cells. Pericellular matrix assembly Excess of proteroglycan monomer and hyaluronan added to cells when microscopically observed Is done. Matrix assembly around trabecular meshwork cells is responsible for aqueous humor outflow As an important component of resistance, the adjustment of this matrix will determine the intraocular pressure. Can be a fixed factor.   1 × 10 5 HTM cells per plate^FourPlated on 35 mm dishes at cell density. cell Was added to DMEM containing 2 mM glutamine, 1 nM penicillin, and 1 nM streptomycin. −H 16/10^-7MT_ThreeCultured for 2 or 6 days with or without. Also, this medium Is a mixed cation / anion exchange resin (AG 501-X8 Resin, Bio-Rad, (Hercules, CA) Hormone removed by incubation with Consisted of 0% fetal calf serum. For exogenous matrix assembly Next, cells were harvested at 3.0 mg / ml (purified from rat chondrosarcoma (Reference 1)). Compatible proteloglycan monomer and 15 μg / ml hyaluronan (grade 1, S igma Chemical Co., St. (Louis, MO) at 37 ° C for 3 hours. The medium is removed and the phosphate buffer containing 0.1% calf serum albumin Formaldehyde-fixed red blood cell suspension in buffered saline (1 x 10⁸cell / 5 ml) was added to each well of cells. After 10 minutes, the cells and extracellular matrix (ECMs) were visualized and observed with a phase contrast microscope (References 2 and 3). these Of cell matrices were classified into one of three groups. With coasts No cells have no visible ECM. A cell with a small coast is It has a visible ECM that extends out of the plasma membrane and less than the width of the cell's nucleus. Big course The cell with the cell moves out of the cell's plasma membrane. It has a visible ECM that extends beyond the width of the cell's nucleus.   Ten^-7MT_ThreeCells cultured for 2 days in the presence of_ThreeCultured in the absence of It had about twice as many cells without coast than cells did. Ten^-7 M's T_Three4 days after treatment, this difference increased more than 4-fold. Therefore, 10^-7M T_ThreeHTM cells treated with_ThreeCells cultured in a medium that does not contain Binds less HA than assembles and assembles smaller ECMs. Therefore , T_ThreeCan act to displace ECM from trabecular cells, or , Can inhibit ECM assembly synthesis. In any case, Decreased vesicle-bound ECM reduces aqueous humor outflow resistance and reduces intraocular pressure Can be made.   Example 12-Synthesis of STHs   Many TR (thyroid receptor) ligands, T4 (thyroxine), T3, Those that include T2 and TS-9 are known in the art. Jorgensen, Thyroid  Hormones and Analogs, in 6 Hormonal Proteins and Peptides, Thyroid Horm ones 107-204 (Choh Hao Lied., 1978) (incorporated herein by reference). checking ...   The synthesis of some TR ligands is described below.   Synthesis of TS1, TS2, TS3, TS4, TS5   TS1, TS2, TS3, TS4 and TS5 and all of their analogs are converted to T3 (3,5 , 3'-triiodo-L-thyronine), T4 (thyroxine) and 3,5-diio Mere acylation of the nitrogen atom of any thyronine analog, including dothyronine Can be prepared. TS1 and TS2 set T3 to Ph, respectively._TwoCHCO_TwoNHS (N-hydroxysuccinimide-2,2-diphenylacetate) and C₁₆H₃₃ CO_TwoSynthesized by reacting with NHS. TS3, T3, FMOC- Cl (fluorenylmethyloxycarbonyl chloride) To achieve. TS4, T3, tBOC_TwoO (tBOC anhydride or di-t-butyl dicarbonate G) to synthesize. R_Three ¹Also -H in place of -I TS5 different from TS1-4, and 3,5-diiodothyronine to tBOC_TwoO and anti It is synthesized by reacting. General for TS1, TS2, TS3, TS4 and TS5 The reaction scheme is shown in FIG. In this reaction scheme, TS5 and its precursor Both are R_Three ¹It should be noted that it has hydrogen rather than iodine at the position.   Combination of TS6 and TS7   TS6 is synthesized by reacting TS5 with paranitrophenyl isocyanate. To achieve. TS7 is reacted with TFA (trifluoroacetic acid), which cleaves the tBOC group. To synthesize. These reactions are performed by anyone in the art. Is a simple organic synthesis reaction. Diagram of the synthesis scheme for TS6 and TS7 It is shown in FIG.   Synthesis of TS8   TS8 is converted to triethylamine and any amide-forming condensing agent such as TBTU ( (Hydroxybenztriazole uronium tetrafluoroborate) or HBTU (Hydroxybenztriazole uronium hexafluorophosphate) Ph in TS5 in the presence_TwoCHNH_Two(Diphenylmethylamine) Combine. The synthesis scheme for TS8 is depicted in FIG.   Synthesis of 3,5-diiodo-3'-isopropylthyronine derivative   In order to design a class of antagonists, a 3 ' It is important to have an aqueous group and to have an extension at the 5 'position. That Preferred hydrophobic groups at the 3 'position are: methyl, benzyl, phenyl, iodo, and And heterocyclic structures. 3, The synthesis of 5-diiodo-3'-isopropyl-5'-substituted thyronines is described below. I do. The examples provided describe special steps for synthesizing TS10 compounds Is characterized in that the general reaction scheme has an extension at the 5'-position 3,5-diiodo-3'-isopropyl-5'-substituted thyronine derivatives of any number Can be used by those skilled in the art to synthesize Addition of this class Compounds can also be synthesized using known organic synthesis techniques.   The synthesis of TS10 is described below and is depicted in FIG. Reaction products for each step The numbers used in the reaction scheme for TS10 indicating are in parentheses.   2-formyl-6-isopropylanisole (1): 2-formyl-6-iso Propylanisole (10.0 g, 61 mmol), Casiraghi, et al. JCS Perkin I, 18 62 (1980) (incorporated by reference) was placed in a 50 ml round bottom flask. Added dropwise to a suspension of sodium hydride (3.7 g, 153 mmol) in mL THF and 50 mL DMF . This addition creates an exothermic reaction and the formation of a gray solid. Next, methyl iodide (26. 0 g, 183 mmol) are added dropwise and the reaction mixture is stirred for 5 hours at room temperature. This anti The reaction mixture is quenched with 20 mL of water, then poured into 500 mL of water and treated with ether ( 2 × 300 mL). The ether layers were combined, washed with water (5 × 1000 mL), Dry over magnesium sulfate and concentrate in vacuo to give 10.2 g (94%) of the table The title compound is obtained. below With.   2- (2-hydroxynonyl) -6-isopropylanisole ( The scheme is not shown):   Octylmagnesium chloride (8.4 mL, 16.9 mmol, 2.0 M) in 10 mL at -78 ° C Add dropwise to a solution of 1 (1.5 g, 8.4 mmol) in THF. Warm the reaction mixture to room temperature While stirring for 2 hours. The reaction mixture is diluted with 50 mL ether and 50 mL Pour into water. Wash the ether layer with brine (1 x 50 mL) and dry over sodium sulfate Dry and concentrate in vacuo. Flash chromatography (silica Gel, 10% ether / hexane → 15% ether / hexane) to give the title compound 734 mg (30%) are obtained. below Found: 292.2404; calculated: 292.2402.   2-nonyl-6-isopropylanisole (2):   Compound 2 (663 mg, 2.3 mmol) was dissolved in a solution of 5 mL of ethanol and 5 mL of acetic acid, Then a spatula tip of palladium on carbon catalyst is added. Next, Fill the mixture with hydrogen gas (using a simple balloon and needle) and mix this mixture Is stirred overnight at room temperature. The next day, pour the reaction mixture into ether (100 mL) and allow The ether layer is then extracted with saturated sodium bicarbonate (3 × 100 mL). This a The tellurium layer was dried over sodium sulfate and concentrated in vacuo to 581 mg (91 %) Of (2) is obtained. Found: 276.2459; calc: 276.2453.   Thyronine adduct (4):   Fuming nitric acid (0.071 mL) is added to 0.184 mL of acetic anhydride cooled to -5 ° C. You Sodium (66 mg) is added to this mixture followed by trifluoroacetic acid (0.124 mL). The mixture was stirred for 1 hour while warming to room temperature, at which point all of the iodine was Dissolve. The reaction mixture is then concentrated in vacuo to provide an oily semi-solid material. Was. This residue was dissolved in 0.7 mL of acetic anhydride and cooled to -20 ° C. 1.2mL A solution of anisole (2) (581 mg, 2.1 mmol) in acetic anhydride and 0.58 mL TFA is added dropwise. I do. The reaction mixture was stirred for 1 hour at -20 ° C, then stirred overnight while warming to room temperature. Mix. The reaction mixture is partitioned between water and methylene chloride. This methyl chloride The ren layer is dried over sodium sulfate and concentrated in vacuo to give iodine as an oil. A donium salt (3) is obtained. This material is not purified or characterized and Introduced directly during the coupling reaction.   N. Lewis and P.S. Wallbank, Synthesis 1103 (1987) (incorporated by citation) N-trifluoroacetyl-3,5-diiodosilo prepared according to the procedure of Nin methyl ester (552 mg, 1.0 mmol) and crude iodonium salt from above ( Dissolve all of 3) in 5 mL of anhydrous methanol. Diazabicyclo [5.4. 0] Undecane (DBU) (183mg, 1.2mmol) and copper-bronze with spatula tip Is added and the resulting mixture is stirred overnight at room temperature. The next day, the reaction mixture Is filtered and the filtrate is concentrated in vacuo. Flash chromatography of crude residue 30 mg purified by chromatography (silica gel, 10% ethyl acetate / hexane) (4%) of the protected thyronine adduct (4) is obtained.   Deprotected thyronine (TS10):2.2 protected thyronine 4 (30 mg, 0.04 mmol) Dissolve in a mixture of 5 mL acetic acid and 2.25 mL 49% hydrobromic acid. Allow the reaction mixture to run for 5 hours Heat to reflux. This reaction mixture is Cool to room temperature and remove these solvents in vacuo. Water to the above oily residue Add to grind to a gray solid. The solid material is filtered, washed with water, and P in vacuum_TwoO_FiveDry on top to obtain 24 mg (81%) of the title compound, TS10. With.   As described above, this reaction scheme is adapted to 3,5-diiodo-3'-isopropyl To synthesize one class of compounds featuring thyronine derivatives, one skilled in the art Can be modified. Here, (1) the 3 'isopropyl group is methyl, Can be substituted with hydrophobic groups containing benzyl, phenyl, iodo, and heterocyclic structures. And (2) a wide variety of chemical structures, including alkyl groups, planar aryls, An element containing a cyclic group or a polar and / or charged group is incorporated at the 5'-position. Can be   Aldehyde (1) in the above reaction scheme is added to the 5'-position of the final thyronine derivative. Is a versatile synthetic intermediate that allows the attachment of various chemical moieties. Furthermore, A variety of chemical reactions may be used to attach these chemical moieties. it can. These reactions are well known in the art, and Addition of organometallics to aldehydes, including primary reagents, organolithiums, etc. Reductive amination of the aldehyde with a secondary amine, and phosphorus ylide or Includes Wittig olefination reaction with stabilized phosphonate anion No. Another possibility is that a benzylic acid which allows an etherification reaction at its 5 'position. Including the reaction of aldehydes to alcohol. As mentioned above, these methods are Base, flat A wide variety of chemical structures, including aryl, heterocyclic or polar and / or charged groups, are It is allowed to be incorporated at the 5'-position of the terminal thyronine derivative.   Table 1 and FIG. 6 represent the structures of some TR ligands.   Where R₆, R_TwoAnd 'R_ThreeRepresents H in Formula 1 and X represents O.   All publications and patent applications mentioned in this specification are to be read as if they were individual publications or publications. States that patent applications are intended to be specifically and individually incorporated by reference. It is incorporated herein by reference to the same extent.   Although the present invention has been fully described, many changes and modifications will occur to those skilled in the art. Can be made to the invention without departing from the spirit or scope of the appended claims. It is clear that you can do that.

[Procedure amendment] [Submission date] December 24, 1999 (December 24, 1999) [Content of amendment] Claims 1. A method for identifying a compound useful as a therapeutic agent, comprising: contacting a compound with trabecular meshwork cells; and detecting the binding of the compound to the trabecular meshwork cells, wherein the compound is 10 micromolar or less. An identification method characterized by having a concentration of: 2. 2. The method of claim 1, wherein the detecting comprises measuring a decrease in hyaluronic acid secretion from human trabecular meshwork cells, and further comprising selecting a compound useful in modulating glycosaminoglycan production. the method of. 3. 2. The method of claim 1, wherein the detecting step comprises measuring binding of the compound to an intracellular receptor in human trabecular meshwork cells. 4. 3. The method of claim 2, wherein said compound is a synthetic thyroid hormone. 5. 5. The method of claim 4, wherein the synthetic thyroid hormone is not an esterified derivative of a synthetic thyroid hormone. 6. 5. The method of claim 4, wherein said synthetic thyroid hormone is selected from the group consisting of TS1, TS2, TS3, TS4, TS5, TS6, TS7, TS8, TS9 and TS10, or an esterified derivative thereof. 7. 7. The method of claim 6, wherein said synthetic thyroid hormone is not an ester derivative. 8. Further comprising the step of synthesizing the synthetic thyroid hormone, and wherein the concentration of the synthetic thyroid hormone at a concentration of 1.0 micromolar or less causes the hyaluronic acid secretion from the trabecular meshwork cells cultured in the absence of the synthetic thyroid hormone. 5. The method of claim 4, wherein said method reduces hyaluronic acid secretion by at least 30%. 9. 2. The method of claim 1, further comprising the step of topically administering said synthetic thyroid hormone to a mammalian eye except a human, and wherein said trabecular meshwork cells are isolated from a rabbit, rat, mouse, pig, cat or monkey eye. The method described in. Ten. The detecting step further comprises indirectly measuring the binding of the compound in the presence and absence of the compound using a functional assay for measuring the binding of the compound to the trabecular meshwork cells. The method of claim 1, comprising: 11． A pharmaceutical composition for inhibiting hyaluronic acid secretion, comprising a synthetic thyroid hormone that inhibits hyaluronic acid secretion from cells that secrete hyaluronic acid, wherein the synthetic thyroid hormone has a concentration of 10 micromolar or less, At least 10% of the total hyaluronic acid secretion from cultured human trabecular meshwork cells cultured at 37 ° C for 3 days is compared to human trabecular meshwork cells cultured at 37 ° C for 3 days in the absence of the synthetic thyroid hormone. A pharmaceutical composition characterized by inhibiting. 12． 12. The pharmaceutical composition according to claim 11, wherein said synthetic thyroid hormone inhibits at least 10% of total hyaluronic acid secretion from cultured human trabecular meshwork cells at a concentration of 1 micromolar. 13. 13. The pharmaceutical composition according to claim 12, wherein said synthetic thyroid hormone inhibits at least 10% of total hyaluronic acid secretion from cultured human trabecular meshwork cells at a concentration of 0.1 micromolar. 14. 13. The pharmaceutical composition according to claim 12, wherein said cells are human trabecular meshwork cells. 15． 15. The pharmaceutical composition according to claim 14, wherein said human trabecular meshwork cells are in the trabecular meshwork. 16． 15. The pharmaceutical composition according to claim 14, wherein said pharmaceutical composition further comprises an ophthalmically compatible pharmaceutical carrier. 17． 17. The pharmaceutical composition according to claim 16, wherein said pharmaceutical carrier is an eye-compatible physiological saline. 18． 17. The pharmaceutical composition according to claim 16, wherein said synthetic thyroid hormone is an ester derivative. 19. 17. The pharmaceutical composition according to claim 16, wherein said synthetic thyroid hormone is placed in a liposome mixture. 20. 17. The pharmaceutical composition according to claim 16, wherein the synthetic thyroid hormone is placed in a synthetic polymer that can be released by diffusion. twenty one. A pharmaceutical composition for the treatment of glaucoma, comprising an ophthalmologically effective amount of a synthetic thyroid hormone, provided that it is not used for the treatment of glaucoma in eyes having cataract. twenty two. 22. The pharmaceutical composition according to claim 21, further comprising an ophthalmically compatible pharmaceutical carrier. twenty three. 23. The pharmaceutical composition according to claim 22, wherein said composition is incorporated into an ocular insert or implant. twenty four. 24. The pharmaceutical composition of claim 23, wherein the ocular implant is a contact impregnated with the synthetic thyroid hormone in preparation for diffusion of the synthetic thyroid hormone from the contact. twenty five. 23. The pharmaceutical composition of claim 22, wherein the ophthalmically compatible pharmaceutical carrier comprises a biodegradable polymer capable of sustained release of the synthetic thyroid hormone. 26. 22. The pharmaceutical composition according to claim 21, wherein said effective amount is 0.1 ng to 20 ng / one eye / day. 27. At a concentration of the synthetic thyroid hormone of 1 micromolar or less, the hyaluronic acid secretion from the trabecular meshwork cells is less than that of the trabecular meshwork cells cultured in the absence of the synthetic thyroid hormone. 26. The pharmaceutical composition according to claim 25, which reduces by at least 30%. 28. 22. The pharmaceutical composition of claim 21, wherein the composition is included in an intraocular sustained release device or surgical implant comprising a polymer capable of sustained release of the synthetic thyroid hormone. 29. 29. The pharmaceutical composition according to claim 28, wherein said polymer is biodegradable. 30. A pharmaceutical composition comprising synthetic thyroid hormone in trabecular meshwork cells, wherein said synthetic thyroid hormone does not cross the blood-eye barrier. 31. 31. The pharmaceutical composition according to claim 30, wherein said synthetic thyroid hormone is an esterified derivative of a synthetic thyroid hormone. 32. 31. The pharmaceutical composition of claim 30, wherein the trabecular meshwork cells are human trabecular meshwork cells from an ocular explant. 33. 31. The pharmaceutical composition according to claim 30, wherein said synthetic thyroid hormone is bound to a human thyroid receptor. 34. 34. The pharmaceutical composition of claim 33, wherein the synthetic thyroid hormone has a trabecular meshwork cell concentration of at least 0.04 ng / dl, and wherein the trabecular meshwork cells are human trabecular meshwork cells. 35. A pharmaceutical composition comprising a synthetic thyroid hormone and an ophthalmically compatible pharmaceutical carrier. 36. 36. The pharmaceutical composition according to claim 35, wherein the synthetic thyroid hormone is an esterified derivative of a synthetic thyroid hormone. 37. 36. The pharmaceutical composition of claim 35, wherein said ophthalmically compatible pharmaceutical carrier comprises a buffered saline solution. 38. 38. The pharmaceutical composition of claim 37, wherein said ophthalmically compatible pharmaceutical carrier comprises a liposome. 39. 36. The pharmaceutical composition of claim 35, wherein said ophthalmically compatible pharmaceutical carrier is a biodegradable synthetic polymer. 40. A pharmaceutical composition comprising an ophthalmic contact impregnated with synthetic thyroid hormone. 41. 41. The pharmaceutical composition according to claim 40, wherein said synthetic thyroid hormone is an esterified derivative of a synthetic thyroid hormone. 42. A pharmaceutical composition for the treatment of glaucoma, comprising an effective amount of a synthetic thyroid hormone. 43. A kit for screening synthetic thyroid hormone for effects on aqueous humor outflow and reduction of intraocular pressure, comprising: a) synthetic thyroid hormone for contacting trabecular meshwork cells at a concentration of 10 micromolar or less; and b) A kit comprising means for measuring hydraulic conductivity of said trabecular meshwork cells after contact with said synthetic thyroid hormone. 44. The synthetic thyroid hormone has the following formula I: Where R ₁ is R ₂ is —H, halogen, CF ₃ , OH, NH ₂ , SH, CH ₃ or —Et, and R ₃ is —H, halogen, —CF ₃ , —OH, —NH ₂ , —N _3, -SH, a -CH ₃ or -Et, R ₅ is -H, _{halogen, -CF 3, -OH, -NH 2} , -N 3, -SH, a -CH ₃ or -Et, R ₆ is -H, halogen, -CF ₃ , -OH, -NH ₂ , -SH or -CH ₃ , and R ' ₂ is -H, halogen, -CF ₃ , -OH, -NH ₂ , -N ₃ , -SH, a -CH ₃ or -Et, R _'3 is -H, halogen, -CF _3, -SH, alkyl, aryl, 5-membered or 6-membered heterocyclic aromatic group, or a cyano , R _'4 is a by -H, _{halogen, -CF 3, -OH, -NH 2} , -SH, -CH 3, -Et, alkyl, aryl, or 5 or 6 membered heterocyclic aromatic group 'heterocyclic aromatic group attached _fourth in the O or N, or S, R' R through urea or carbamate linkage Te ₅ is alkyl, aryl, 5- or 6-membered heterocyclic Wherein an aromatic group, a heterocyclic alkyl, heterocyclic aryl, arylalkyl, heterocyclic aryl alkyl, polyaromatic group or a multi-heterocyclic aromatic radical, by the R _'5 is a polar group or charged groups may be substituted, R _'6 is -H, _{halogen, -CF 3, -OH, -NH 2} , -SH, a -CH ₃ or -Et, X is _{O, S, SO 2, NH} , NR ₇ is CH _2, CHR ₇ or CR ₇ R _7, wherein R ₇ is an alkyl, aryl or 5- or 6-membered heterocyclic aromatic group, and 'R ₁ -'R ₆ group and R ₁ any one of -R ₆ group can replace them by binding to B) is defined by the kit according to claim 43. 45. The synthetic thyroid hormone has the following formula II: 44. The kit of claim 43, wherein Y is a group compatible with the ester. 46. A kit for assaying the effect of synthetic thyroid hormone on extracellular matrix construction, comprising: a) a synthetic thyroid hormone for contacting trabecular meshwork cells at a concentration of 10 micromolar or less in the presence of hyaluronic acid and proteoglycan monomers. And b) a kit comprising means for measuring the amount of extracellular matrix bound to said trabecular meshwork cells. 47. The synthetic thyroid hormone has the following formula I: Where R ₁ is R ₂ is —H, halogen, CF ₃ , OH, NH ₂ , SH, CH ₃ or —Et, and R ₃ is —H, halogen, —CF ₃ , —OH, —NH ₂ , —N _3, -SH, a -CH ₃ or -Et, R ₅ is -H, _{halogen, -CF 3, -OH, -NH 2} , -N 3, -SH, a -CH ₃ or -Et, R ₆ is -H, halogen, -CF ₃ , -OH, -NH ₂ , -SH or -CH ₃ , and R ' ₂ is -H, halogen, -CF ₃ , -OH, -NH ₂ , -N ₃ , -SH, a -CH ₃ or -Et, R _'3 is -H, halogen, -CF _3, -SH, alkyl, aryl, 5-membered or 6-membered heterocyclic aromatic group, or a cyano , R _'4 is a by -H, _{halogen, -CF 3, -OH, -NH 2} , -SH, -CH 3, -Et, alkyl, aryl, or 5 or 6 membered heterocyclic aromatic group 'heterocyclic aromatic group attached _fourth in the O or N, or S, R' R through urea or carbamate linkage Te ₅ is alkyl, aryl, 5- or 6-membered heterocyclic Wherein an aromatic group, a heterocyclic alkyl, heterocyclic aryl, arylalkyl, heterocyclic aryl alkyl, polyaromatic group or a multi-heterocyclic aromatic radical, by the R _'5 is a polar group or charged groups may be substituted, R _'6 is -H, _{halogen, -CF 3, -OH, -NH 2} , -SH, a -CH ₃ or -Et, X is _{O, S, SO 2, NH} , NR ₇ is CH _2, CHR ₇ or CR ₇ R _7, wherein R ₇ is an alkyl, aryl or 5- or 6-membered heterocyclic aromatic group, and 'R ₁ -'R ₆ group and R ₁ any one of -R ₆ group can replace them by binding to B) is defined by the kit according to claim 46. 48. The synthetic thyroid hormone has the following formula II: 47. The kit of claim 46, wherein Y is a group compatible with an ester. 49. A pharmaceutical composition for lowering intraocular pressure, comprising a therapeutically effective amount of a synthetic thyroid hormone. 50. 50. The pharmaceutical composition according to claim 49, comprising a pharmaceutically acceptable carrier. 51. The pharmaceutical composition according to claim 50, which is for topical administration. 52. 51. The pharmaceutical composition according to claim 50, for intraocular administration. [Procedure amendment] [Date of submission] January 17, 2000 (2000.1.17) [Contents of amendment] “Fig. 3” on page 51, line 7 of the specification will be amended to “Fig. 5”. "Figure 4" on page 51, 15th line of the specification is corrected to "Figure 6". "Figure 5" on the sixth line from the bottom of page 51 of the specification is corrected to "Figure 7". "Fig. 6" on page 52, line 9 of the specification is corrected to "Fig. 8". "Figure 6" on page 56, 4th line of the specification is corrected to "Figure 9".

────────────────────────────────────────────────── ─── Continuation of front page    (81) Designated countries EP (AT, BE, CH, DE, DK, ES, FI, FR, GB, GR, IE, IT, L U, MC, NL, PT, SE), OA (BF, BJ, CF) , CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AP (GH, KE, LS, MW, S D, SZ, UG, ZW), EA (AM, AZ, BY, KG) , KZ, MD, RU, TJ, TM), AL, AM, AT , AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, F I, GB, GE, GH, HU, IL, IS, JP, KE , KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, M X, NO, NZ, PL, PT, RO, RU, SD, SE , SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, UZ, VN, YU, ZW (72) Inventors Baxter, John Dee.             United States, California 94127,             San Francisco, San Pablo Aveni             Fit 131 (72) Inventor Jumper, Michael Dee.             North Carolina, United States             27705, Durham, Brown Avenue             2729 (72) Inventor Scanlan, Thomas S.             United States of America, California 94122,             Moraga Street, San Francisco             2525

Claims (1)

[Claims] 1. A method of identifying a compound useful as a therapeutic agent, comprising: contacting a compound with trabecular meshwork cells; and detecting binding of the compound to the trabecular meshwork cells, wherein the compound comprises 10 The above-described identification method, wherein the concentration is less than or equal to micromolar. 2. 2. The method of claim 1, wherein said detecting step comprises measuring a decrease in hyaluronic acid secretion from human trabecular meshwork cells, and further comprising selecting a compound useful for modulating GAG production. . 3. 2. The method of claim 1, wherein the detecting step comprises measuring binding of the compound to an intracellular receptor in human trabecular meshwork cells. 4. 3. The method of claim 2, wherein said compound is a synthetic thyroid hormone. 5. 5. The method of claim 4, wherein said synthetic thyroid hormone is an esterified derivative of a synthetic thyroid hormone. 6. The method of claim 4, wherein the synthetic thyroid hormone is selected from the group consisting of TS1, TS2, TS3, TS4, TS5, TS6, TS7, TS8, TS9 and TS10, or an esterified derivative thereof. 7. 7. The method of claim 6, wherein said synthetic thyroid hormone is an ester derivative. 8. Further comprising the step of synthesizing the synthetic thyroid hormone, wherein the synthetic thyroid hormone at a concentration of 1.0 micromolar or less is capable of secreting the hyaluronic acid from the trabecular meshwork cells cultured in the absence of the synthetic thyroid hormone. 5. The method of claim 4, wherein said secretion decreases hyaluronic acid secretion by at least 30%. 9. 2. The method of claim 1, further comprising topically applying the synthetic thyroid hormone to a mammalian eye, and wherein the trabecular meshwork cells have been isolated from a rabbit, rat, mouse, pig, cat or monkey eye. the method of. Ten. The detection step further comprises indirectly measuring the binding of the compound in the presence and absence of the compound by a functional assay for measuring the binding of the compound to the trabecular meshwork cells, The method of claim 1. 11． A method of inhibiting hyaluronic acid secretion, comprising: non-systemically applying a fixed amount of a synthetic thyroid hormone that inhibits hyaluronic acid secretion from cells that secrete hyaluronic acid, wherein the concentration is less than 10 micromolar. The cultured human trabecular meshwork cells cultured at 37 ° C for 3 days as compared to the human trabecular meshwork cells cultured in the absence of the synthetic thyroid hormone after cultured at 37 ° C for 3 days. inhibits at least 10% of the secreted total hyaluronic acid from with the proviso that the non-systemic application are, 1) subconjunctival injection of T _3, or 2) local of T ₃ or T ₄ isolated from native tissue Said method, without application. 12. The method of claim 11, wherein the synthetic thyroid hormone at a concentration of 12.1 micromolar inhibits at least 10% of total hyaluronic acid secretion in cultured human trabecular meshwork cells. 13. The method of claim 12, wherein said synthetic thyroid hormone at a concentration of 0.1 micromolar inhibits at least 10% of total hyaluronic acid secretion in cultured human trabecular meshwork cells. 14. 13. The method of claim 12, wherein said cells are human trabecular meshwork cells. 15． 15. The method of claim 14, wherein said human trabecular meshwork cells are within trabecular meshwork. 16． 15. The method of claim 14, wherein the synthetic thyroid hormone is applied with an ophthalmically compatible pharmaceutical carrier. 17． 17. The method of claim 16, wherein the pharmaceutical carrier is an ophthalmically compatible saline. 18． 17. The method of claim 16, wherein said synthetic thyroid hormone is an ester derivative. 19. 17. The method of claim 16, wherein said synthetic thyroid hormone is in a liposome mixture. 20. 17. The method of claim 16, wherein said synthetic thyroid hormone is disposed in a synthetic polymer capable of releasing said synthetic polymer by diffusion. twenty one. Administering glaucoma to an eye in need of such treatment, wherein the dose is 1) T ₃ or T ₄ conjunctiva. lower injection, 2) topical application of T ₃ or T ₄ isolated from natural tissue, or 3) administration of T ₃ or T ₄ to the eye suffering from cataracts, not including, the methods. twenty two. 22. The method of claim 21, wherein said administering comprises topically applying said synthetic thyroid hormone to the eye in an ophthalmically compatible pharmaceutical carrier. twenty three. 23. The method of claim 22, wherein said topical application comprises diffusion of said synthetic thyroid hormone from an ocular insert or implant. twenty four. 24. The method of claim 23, wherein the ocular implant is an ocular contact impregnated with the synthetic thyroid hormone to allow diffusion of the synthetic thyroid hormone from the contact. twenty five. 23. The method of claim 22, wherein said topical application comprises applying said synthetic thyroid hormone with an ophthalmically compatible pharmaceutical carrier. 26. 26. The method of claim 25, wherein said ophthalmically compatible pharmaceutical carrier comprises a biodegradable polymer capable of sustained release of said STH. 27. 27. The method of claim 26, wherein said effective amount is between 0.1 ng and 20 ng / eye / day. 28. By applying the synthetic thyroid hormone locally to the mammalian eye and measuring the intraocular pressure before and after the topical application step, the synthetic thyroid hormone at a concentration of 1 micromolar or less is converted to a non-molar form of the synthetic thyroid hormone. 27. The method of claim 26, wherein the method reduces hyaluronic acid secretion from the trabecular meshwork cells by at least 30% compared to hyaluronic acid secretion from the trabecular meshwork cells cultured in the presence. 29. 22. The method of claim 21, wherein said administering comprises surgically implanting or injecting an intraocular sustained release device comprising a polymer capable of sustained release of the STH. 30. 30. The method of claim 29, wherein said polymer is biodegradable. 31. A composition comprising synthetic thyroid hormone in trabecular meshwork cells; wherein the synthetic thyroid hormone does not cross the blood-eye barrier. 32. 32. The composition of claim 31, wherein said synthetic thyroid hormone is an esterified derivative of a synthetic thyroid hormone. 33. 32. The composition of claim 31, wherein the trabecular meshwork cells are human trabecular meshwork cells from an ocular explant. 34. 32. The composition of claim 31, wherein said synthetic thyroid hormone binds to a human thyroid receptor. 35. 35. The composition of claim 34, wherein the synthetic thyroid hormone has a concentration in the trabecular meshwork cell of at least 0.04 ng / dl, and wherein the trabecular meshwork cell is a human trabecular meshwork cell. 36. A composition comprising a synthetic thyroid hormone and an ophthalmically compatible pharmaceutical carrier. 37. 37. The composition of claim 36, wherein the synthetic thyroid hormone is an esterified derivative of a synthetic thyroid hormone. 38. 37. The composition of claim 36, wherein said ophthalmically compatible pharmaceutical carrier is a buffered saline solution. 39. 39. The composition of claim 38, wherein said ophthalmically compatible pharmaceutical carrier is a liposome. 40. 39. The composition of claim 36, wherein said ophthalmically compatible pharmaceutical carrier is a biodegradable synthetic polymer. 41. A composition comprising an ocular contact impregnated with a synthetic thyroid hormone. 42. 42. The composition of claim 41, wherein said synthetic thyroid hormone is an esterified derivative of a synthetic thyroid hormone. 43. A method for treating glaucoma, comprising: administering an effective amount of a synthetic thyroid hormone systemically to a patient in need thereof. 44. A method of screening STH for effects on aqueous humor outflow and IOP reduction, comprising: a) contacting trabecular meshwork cells with said STH at a concentration of 10 micromolar or less; b) contacting said small cells with said STH after said contact with said STH. Measuring the hydraulic conductivity of the trabecular meshwork cells. 45. 45. The method of claim 44, wherein said STH is defined by formula (I). 46. 45. The method of claim 44, wherein said STH is defined by formula (II). 47. A method for assaying the effect of STH on extracellular matrix assembly, comprising: a) contacting STH with trabecular meshwork cells at a concentration of 10 micromolar or less in the presence of HA and a proteoglycan monomer; b) the trabecular meshwork Measuring the amount of extracellular matrix bound to the cells. 48. 50. The method of claim 47, wherein said compound is defined by formula (I). 49. 50. The method of claim 47, wherein said compound is defined by formula (II). 50. Use of a known thyroid hormone in a therapeutically effective amount in a composition for reducing intraocular pressure. 51. 51. The method of claim 50, wherein said composition comprises a pharmaceutically acceptable carrier. 52. 52. The method of claim 51, wherein the composition is administered topically. 53. 52. The method of claim 51, wherein said composition is administered intraocularly.